Thiolactams and uses thereof

ABSTRACT

This invention provides compounds of formula I: 
     
       
         
         
             
             
         
       
         
         
           
             wherein R 1 , R 2 , R 3 , R 4 , R 5 , and R 6  are as described in the specification. The compounds are inhibitors of PLK and are thus useful for treating proliferative, inflammatory, or cardiovascular disorders.

The present application is a continuation of U.S. patent applicationSer. No. 12/631,144, filed Dec. 4, 2009 now U.S. Pat. No. 7,998,952,which claims the benefit of U.S. Provisional Application Ser. No.61/200,945, filed Dec. 5, 2008 (abandoned). The entire contents of eachof the above-referenced patent applications are incorporated herein bythis reference.

BACKGROUND OF THE INVENTION

Protein kinases constitute a large family of structurally relatedenzymes that effect the transfer of a phosphate group from a nucleosidetriphosphate to a protein acceptor. A vast array of cellular functions,including DNA replication, cell cycle progression, energy metabolism,and cell growth and differentiation, are regulated by reversible proteinphosphorylation events mediated by protein kinases. Additionally,protein kinase activity has been implicated in a number of diseasestates. Accordingly, protein kinase targets have attracted substantialdrug discovery efforts in recent years, with several protein kinaseinhibitors achieving regulatory approval (reviewed in Fischer, Curr.Med. Chem., 11:1563 (2004); Dancey and Sausville, Nature Rev. DrugDisc., 2:296 (2003)).

PLK is a serine/threonine protein kinase that plays a key role in cellcycle control. PLK controls entry and progression through mitosis atmultiple stages by regulating centrosome maturation, activation ofinitiating factors, degradation of inhibitory components, chromosomecondensation, and exit from mitosis (reviewed in Barr et al., NatureReviews Mol Cell Biol., 5; 429 (2004); Petronczki et al., Dev. Cell, 5;646 (2008)). PLK has been reported to be overexpressed in numerouscancers such as melanoma, prostate, ovarian, colorectal, pancreatic, nonsmall cell lung, esophageal, endometrial, glioma, squamous cellcarcinoma of the head and neck and non-Hodkins lymphoma (Kneisel et al.,J Cutan Pathol 29: 354 (2002); Takai et al. Cancer Lett 169: 41 (2001);Takahashi et al Cancer Sci.; 94(2):148 (2003); Macmillan et al, Ann.Surg. Oncol. 8: 729 (2001); Gray et al. Mol. Cancer. Ther. 3: 641(2004); Dietzmann et al. J. Neurooncol. 53:1 (2001); Ito et al. Br. J.Cancer 90:414 (2005); Weichert et al. Pancreatology. 5:259 (2005); Mitoet al. Leuk. Lymphoma 46:225 (2005); Liu et al. Oncology 74:96 (2008);Yamamoto et al. Oncology 70(3):231 (2006); Weichert et al. Cancer Sci.97(4):271 (2006)). Increased levels of expression are additionallycorrelated with poor prognosis and survival. (Takai et al. Cancer Lett164: 41 (2001); Wolf et al. Oncogene 14: 543 (1997); Knecht et al.Cancer Res. 59 (1999); Strebhardt et al. JAMA 283:479 (2000); Weichertet al. World J. Gastroenterol 11:5644 (2005); Tokumitsu et al. Int. J.Oncol. 15: 687 (1999); Takai et al. Cancer Lett. 164:41 (2001); Weichertet al. Prostate 60:240 (2004); Kanaji et al. Oncology 70(2):126 (2006)).Overexpression of the kinase transforms cells, rendering them oncogenicsuch that they acquire the ability to form tumors in mice (Smith et al.,Biochem. Biophys. Res. Commun. 234; 397 (1997)). PLK protein levels arealso elevated in tumor relative to normal cell lines in culture.Downregulation of PLK protein expression by RNA interference in tumorcell lines results in a reduction of cell proliferation, mitotic arrestat prometaphase and the rapid progression into apoptosis(Spankuch-Schmitt et al. J. Natl. Cancer Inst. 94(24):1863 (2002);Spankuch-Schmitt et al. Oncogene 21(20):3162 (2002)). This effect wasnot observed in normal cell lines. Moreover downregulation of PLK byshort hairpin expression in mice with human xenografts reduced tumorgrowth to 18% (Spankuch et al. J. Natl. Cancer Inst. 96(11):862 (2004);Kappel et al. Nucleic Acids Res. 34(16) 4527 (2006)). The key role ofPLK in mitotic progression, its overexpression in a wide range ofmalignancies and the anti-proliferative effect observed upon itsinhibition demonstrate its feasibility as a therapeutic target.

Accordingly, inhibitors of PLK are useful for treating various diseasesor conditions associated with PLK activity, and are especially needed inview of the inadequate treatments currently available for many of thesedisorders.

DETAILED DESCRIPTION OF THE INVENTION

1. General Description of Compounds of the Invention:

This invention provides compounds that are inhibitors of PLK, andaccordingly are useful for the treatment of proliferative, inflammatory,or cardiovascular disorders. The compounds of this invention arerepresented by formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is selected from hydrogen, —CN, halogen, optionally substitutedC₁₋₆aliphatic, or —YR^(1a),

-   -   wherein Y is —O—, —S—, or —NR^(1a), and each occurrence of        R^(1a) is independently hydrogen, or optionally substituted        C₁₋₆aliphatic;

R² is selected from hydrogen, halogen, —ZR^(2a), or —OR^(2b),

-   -   wherein Z is an optionally substituted C₁₋₆ alkylene chain, and        R^(2a) is —OR^(2b), —N(R^(2b))₂, —SR^(2b), —C(O)N(R^(2b))₂,        —N(R^(2b))C(O)R^(2b), —SO₂N(R^(2b))₂, —NR^(2b)SO₂R^(2b),        —NR^(2b)C(O)N(R^(2b))₂, or —NR^(2b)SO₂N(R^(2b))₂, wherein each        occurrence of R^(2b) is independently hydrogen or optionally        substituted C₁₋₆alkyl, or two occurrences of R^(2b), taken        together with a nitrogen atom to which they are bound, form an        optionally substituted 3-7-membered heterocyclyl ring;

R³ is selected from hydrogen, halogen, optionally substituted C₁₋₄alkyl,or optionally substituted C₁₋₄alkoxy;

R⁴ is selected from hydrogen, optionally substituted C₁₋₆aliphatic, anoptionally substituted 3-7-membered heterocyclyl ring,—(CH₂)_(x)NR^(4a)R^(4b), —(CH₂)_(x)NR^(4a)C(O)R^(4b),—(CH₂)_(x)NR^(4a)S(O)₂R^(4b), —(CH₂)_(x)C(O)R^(4b),—(CH₂)_(x)C(O)NR^(4a)R^(4b), —(CH₂)_(x)S(O)₂NR^(4a)R^(4b), or—(CH₂)_(x)OR^(4b),

wherein

each occurrence of x is independently 0-6;

wherein R^(4a) is hydrogen or optionally substituted C₁₋₆aliphatic, and

R^(4b) is hydrogen, optionally substituted C₁₋₆aliphatic, optionallysubstituted C₃₋₇heterocyclyl or C₃₋₇carbocyclyl ring, or is W—R^(4c),wherein W is an optionally substituted C₂₋₆ alkylene chain, and R^(4c)is an optionally substituted C₃₋₇-heterocyclyl ring, —OR^(4d),—N(R^(4d))₂, —SR^(4d), —C(O)N(R^(4d))₂, —N(R^(4d))C(O)R^(4d),—SO₂N(R^(4d))₂, —NR^(4d)SO₂R^(4d), —NR^(4d)C(O)N(R^(4d))₂, or—NR^(4d)SO₂N(R^(4d))₂, wherein each occurrence of R^(4d) isindependently hydrogen or optionally substituted C₁₋₆aliphatic, or twooccurrences of R^(4d), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted 3-7-membered heterocyclylring;

or wherein R^(4a) and R^(4b), taken together with the nitrogen atom towhich they are bound, form an optionally substituted 3-7-memberedheterocyclyl ring;

R⁵ is hydrogen, optionally substituted C₁₋₆aliphatic, an optionallysubstituted C₃₋₇₋heterocyclyl ring, or is X—R^(5a), wherein X is anoptionally substituted C₂₋₆ alkylene chain or —NR^(5c),

-   -   wherein when X is an optionally substituted C₂₋₆ alkylene chain        R^(5a) is —OR^(5b), —N(R^(5b))₂, —SR^(5b), —C(O)N(R^(5b))₂,        —N(R^(5b))C(O)R^(5b), —SO₂N(R^(5b))₂, —NR^(5b)SO₂R^(5b),        —NR^(5b)C(O)N(R^(5b))₂, or —NR^(5b)SO₂N(R^(5b))₂; and    -   when X is —NR^(5c), R^(5a) is hydrogen or optionally substituted        C₁₋₆aliphatic, or R^(5a) and R^(5c), taken together with the        nitrogen atom to which they are bound, form an optionally        substituted 3-7-membered heterocyclyl ring;    -   wherein each occurrence of R^(5b) and R^(5c) is independently        hydrogen or optionally substituted C₁₋₆aliphatic, or two        occurrences of R^(5b), or R^(5a) and R^(5c), taken together with        the nitrogen atom to which they are bound, form an optionally        substituted 3-7-membered heterocyclyl ring; or

wherein R⁴ and R⁵, taken together, form an optionally substituted5-7-membered cycloaliphatic or heterocyclyl ring; and

R⁶ is selected from hydrogen, halogen, optionally substituted C₁₋₄alkyl,or optionally substituted C₁₋₄alkoxy.

2. Compounds and Definitions:

Compounds of this invention include those described generally forformula I above, and are further illustrated by the classes, subclasses,and species disclosed herein. As used herein, the following definitionsshall apply unless otherwise indicated.

As described herein, compounds of the invention may be optionallysubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted”,whether preceded by the term “optionally” or not, means that a hydrogenradical of the designated moiety is replaced with the radical of aspecified substituent, provided that the substitution results in astable or chemically feasible compound. The term “substitutable”, whenused in reference to a designated atom, means that attached to the atomis a hydrogen radical, which hydrogen atom can be replaced with theradical of a suitable substituent. Unless otherwise indicated, an“optionally substituted” group may have a substituent at eachsubstitutable position of the group, and when more than one position inany given structure may be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at every position. Combinations of substituents envisionedby this invention are preferably those that result in the formation ofstable or chemically feasible compounds.

A stable compound or chemically feasible compound is one in which thechemical structure is not substantially altered when kept at atemperature from about −80° C. to about +40°, in the absence of moistureor other chemically reactive conditions, for at least a week, or acompound which maintains its integrity long enough to be useful fortherapeutic or prophylactic administration to a patient.

The phrase “one or more substituents”, as used herein, refers to anumber of substituents that equals from one to the maximum number ofsubstituents possible based on the number of available bonding sites,provided that the above conditions of stability and chemical feasibilityare met.

As used herein, the term “independently selected” means that the same ordifferent values may be selected for multiple instances of a givenvariable in a single compound.

As used herein, “a 3-7-membered saturated, partially unsaturated, oraromatic monocyclic ring having 0-3 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur, or an 8-10-membered partiallyunsaturated, or aromatic bicyclic ring system having 0-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur” includescycloaliphatic, heterocyclic, aryl and heteroaryl rings.

As used herein, the term “aromatic” includes aryl and heteroaryl groupsas described generally below and herein.

The term “aliphatic” or “aliphatic group”, as used herein, means anoptionally substituted straight-chain or branched C₁₋₁₂ hydrocarbon, ora cyclic C₁₋₁₂ hydrocarbon which is completely saturated or whichcontains one or more units of unsaturation, but which is not aromatic(also referred to herein as “carbocycle”, “cycloaliphatic”,“cycloalkyl”, or “cycloalkenyl”). For example, suitable aliphatic groupsinclude optionally substituted linear, branched or cyclic alkyl,alkenyl, alkynyl groups and hybrids thereof, such as (cycloalkyl)alkyl,(cycloalkenyl)alkyl, or (cycloalkyl)alkenyl. Unless otherwise specified,in various embodiments, aliphatic groups have 1-12, 1-10, 1-8, 1-6, 1-4,1-3, or 1-2 carbon atoms.

The term “alkyl”, used alone or as part of a larger moiety, refers to anoptionally substituted straight or branched chain hydrocarbon grouphaving 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms.

The term “alkenyl”, used alone or as part of a larger moiety, refers toan optionally substituted straight or branched chain hydrocarbon grouphaving at least one double bond and having 2-12, 2-10, 2-8, 2-6, 2-4, or2-3 carbon atoms.

The term “alkynyl”, used alone or as part of a larger moiety, refers toan optionally substituted straight or branched chain hydrocarbon grouphaving at least one triple bond and having 2-12, 2-10, 2-8, 2-6, 2-4, or2-3 carbon atoms.

The terms “cycloaliphatic”, “carbocycle”, “carbocyclyl”, “carbocyclo”,or “carbocyclic”, used alone or as part of a larger moiety, refer to anoptionally substituted saturated or partially unsaturated cyclicaliphatic ring system having from 3 to about 14 ring carbon atoms. Insome embodiments, the cycloaliphatic group is an optionally substitutedmonocyclic hydrocarbon having 3-8 or 3-6 ring carbon atoms.Cycloaliphatic groups include, without limitation, optionallysubstituted cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl,cyclooctenyl, or cyclooctadienyl. The terms “cycloaliphatic”,“carbocycle”, “carbocyclyl”, “carbocyclo”, or “carbocyclic” also includeoptionally substituted bridged or fused bicyclic rings having 6-12,6-10, or 6-8 ring carbon atoms, wherein any individual ring in thebicyclic system has 3-8 ring carbon atoms.

The term “cycloalkyl” refers to an optionally substituted saturated ringsystem of about 3 to about 10 ring carbon atoms. Exemplary monocycliccycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, and cycloheptyl.

The term “cycloalkenyl” refers to an optionally substituted non-aromaticmonocyclic or multicyclic ring system containing at least onecarbon-carbon double bond and having about 3 to about 10 carbon atoms.Exemplary monocyclic cycloalkenyl rings include cyclopentyl,cyclohexenyl, and cycloheptenyl.

The terms “haloaliphatic”, “haloalkyl”, “haloalkenyl” and “haloalkoxy”refer to an aliphatic, alkyl, alkenyl or alkoxy group, as the case maybe, which is substituted with one or more halogen atoms. As used herein,the term “halogen” or “halo” means F, Cl, Br, or I. The term“fluoroaliphatic” refers to a haloaliphatic wherein the halogen isfluoro, including perfluorinated aliphatic groups. Examples offluoroaliphatic groups include, without limitation, fluoromethyl,difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl,1,1,2-trifluoroethyl, 1,2,2-trifluoroethyl, and pentafluoroethyl.

The term “heteroatom” refers to one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The terms “aryl” and “ar-”, used alone or as part of a larger moiety,e.g., “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refer to an optionallysubstituted C₆₋₁₄aromatic hydrocarbon moiety comprising one to threearomatic rings. Preferably, the aryl group is a C₆₋₁₀aryl group. Arylgroups include, without limitation, optionally substituted phenyl,naphthyl, or anthracenyl. The terms “aryl” and “ar-”, as used herein,also include groups in which an aryl ring is fused to one or morecycloaliphatic rings to form an optionally substituted cyclic structuresuch as a tetrahydronaphthyl, indenyl, or indanyl ring. The term “aryl”may be used interchangeably with the terms “aryl group”, “aryl ring”,and “aromatic ring”.

An “aralkyl” or “arylalkyl” group comprises an aryl group covalentlyattached to an alkyl group, either of which independently is optionallysubstituted. Preferably, the aralkyl group is C₆₋₁₀ arylC₁₋₆alkyl,including, without limitation, benzyl, phenethyl, and naphthylmethyl.

The terms “heteroaryl” and “heteroar-”, used alone or as part of alarger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”, refer togroups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. A heteroarylgroup may be mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, ortricyclic, more preferably mono- or bicyclic. The term “heteroatom”refers to nitrogen, oxygen, or sulfur, and includes any oxidized form ofnitrogen or sulfur, and any quaternized form of a basic nitrogen. Forexample, a nitrogen atom of a heteroaryl may be a basic nitrogen atomand may also be optionally oxidized to the corresponding N-oxide. When aheteroaryl is substituted by a hydroxy group, it also includes itscorresponding tautomer. The terms “heteroaryl” and “heteroar-”, as usedherein, also include groups in which a heteroaromatic ring is fused toone or more aryl, cycloaliphatic, or heterocycloaliphatic rings.Nonlimiting examples of heteroaryl groups include thienyl, furanyl,pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl,pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl,naphthyridinyl, pteridinyl, indolyl, isoindolyl, benzothienyl,benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl,quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl,quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl,phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Theterm “heteroaryl” may be used interchangeably with the terms “heteroarylring”, “heteroaryl group”, or “heteroaromatic”, any of which termsinclude rings that are optionally substituted. The term “heteroaralkyl”refers to an alkyl group substituted by a heteroaryl, wherein the alkyland heteroaryl portions independently are optionally substituted.

As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclicradical”, and “heterocyclic ring” are used interchangeably and refer toa stable 3- to 8-membered monocyclic or 7-10-membered bicyclicheterocyclic moiety that is either saturated or partially unsaturated,and having, in addition to carbon atoms, one or more, preferably one tofour, heteroatoms, as defined above. When used in reference to a ringatom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or NR⁺ (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl,diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl. Aheterocyclyl group may be mono-, bi-, tri-, or polycyclic, preferablymono-, bi-, or tricyclic, more preferably mono- or bicyclic. The term“heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted. Additionally, a heterocyclic ring alsoincludes groups in which the heterocyclic ring is fused to one or morearyl rings.

As used herein, the term “partially unsaturated” refers to a ring moietythat includes at least one double or triple bond between ring atoms. Theterm “partially unsaturated” is intended to encompass rings havingmultiple sites of unsaturation, but is not intended to include aromatic(e.g., aryl or heteroaryl) moieties, as herein defined.

The term “alkylene” refers to a bivalent alkyl group. An “alkylenechain” is a polymethylene group, i.e., —(CH₂)_(n)—, wherein n is apositive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from1 to 2, or from 2 to 3. An optionally substituted alkylene chain is apolymethylene group in which one or more methylene hydrogen atoms isoptionally replaced with a substituent. Suitable substituents includethose described below for a substituted aliphatic group and also includethose described in the specification herein. It will be appreciated thattwo substituents of the alkylene group may be taken together to form aring system. In certain embodiments, two substituents can be takentogether to form a 3-7-membered ring. The substituents can be on thesame or different atoms.

An alkylene chain also can be optionally interrupted by a functionalgroup. An alkylene chain is “interrupted” by a functional group when aninternal methylene unit is interrupted by the functional group. Examplesof suitable “interrupting functional groups” are described in thespecification and claims herein.

For purposes of clarity, all bivalent groups described herein,including, e.g., the alkylene chain linkers described above, areintended to be read from left to right, with a correspondingleft-to-right reading of the formula or structure in which the variableappears.

An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) orheteroaryl (including heteroaralkyl and heteroarylalkoxy and the like)group may contain one or more substituents and thus may be “optionallysubstituted”. In addition to the substituents defined above and herein,suitable substituents on the unsaturated carbon atom of an aryl orheteroaryl group also include and are generally selected from -halo,—NO₂, —CN, —R⁺, —C(R⁺)═C(R⁺)₂, —C≡C—R⁺, —OR⁺, —SR^(o), —S(O)R^(o),—SO₂R^(o), —SO₃R⁺, —SO₂N(R⁺)₂, —N(R⁺)₂, —NR⁺C(O)R⁺, —NR⁺C(S)R⁺,—NR⁺C(O)N(R⁺)₂, —NR⁺C(S)N(R⁺)₂, —N(R⁺)C(═NR⁺)—N(R⁺)₂,—N(R⁺)C(═NR⁺)—R^(o), —NR⁺CO₂R⁺, —NR⁺SO₂R^(o), —NR⁺SO₂N(R⁺)₂, —O—C(O)R⁺,—O—CO₂R⁺, —OC(O)N(R⁺)₂, —C(O)R⁺, —C(S)R^(o), —CO₂R⁺, —C(O)—C(O)R⁺,—C(O)N(R⁺)₂, —C(S)N(R⁺)₂, —C(O)N(R⁺)—OR⁺, —C(O)N(R⁺)C(═NR⁺)—N(R⁺)₂,—N(R⁺)C(═NR⁺)—N(R⁺)—C(O)R⁺, —C(═NR⁺)—N(R⁺)₂, —C(═NR⁺)—OR⁺,—N(R⁺)—N(R⁺)₂, —C(═NR⁺)—N(R⁺)—OR⁺, —C(R^(o))═N—OR⁺, —P(O)(R⁺)₂,—P(O)(OR⁺)₂, —O—P(O)—OR⁺, and —P(O)(NR⁺)—N(R⁺)₂, wherein R⁺,independently, is hydrogen or an optionally substituted aliphatic, aryl,heteroaryl, cycloaliphatic, or heterocyclyl group, or two independentoccurrences of R⁺ are taken together with their intervening atom(s) toform an optionally substituted 5-7-membered aryl, heteroaryl,cycloaliphatic, or heterocyclyl ring. Each R^(o) is an optionallysubstituted aliphatic, aryl, heteroaryl, cycloaliphatic, or heterocyclylgroup.

An aliphatic or heteroaliphatic group, or a non-aromatic carbycyclic orheterocyclic ring may contain one or more substituents and thus may be“optionally substituted”. Unless otherwise defined above and herein,suitable substituents on the saturated carbon of an aliphatic orheteroaliphatic group, or of a non-aromatic carbocyclic or heterocyclicring are selected from those listed above for the unsaturated carbon ofan aryl or heteroaryl group and additionally include the following: ═O,═S, ═C(R*)₂, ═N—N(R*)₂, ═N—OR*, ═N—NHC(O)R*, ═N—NHCO₂R^(o)═N—NHSO₂R^(o)or ═N—R* where R^(o) is defined above, and each R* is independentlyselected from hydrogen or an optionally substituted C₁₋₆ aliphaticgroup.

In addition to the substituents defined above and herein, optionalsubstituents on the nitrogen of a non-aromatic heterocyclic ring alsoinclude and are generally selected from —R⁺, —N(R⁺)₂, —C(O)R⁺, —C(O)OR⁺,—C(O)C(O)R⁺, —C(O)CH₂C(O)R⁺, —S(O)₂R⁺, —S(O)₂N(R⁺)₂, —C(S)N(R⁺)₂,—C(═NH)—N(R⁺)₂, or —N(R⁺)S(O)₂R⁺; wherein each R⁺ is defined above. Aring nitrogen atom of a heteroaryl or non-aromatic heterocyclic ringalso may be oxidized to form the corresponding N-hydroxy or N-oxidecompound. A nonlimiting example of such a heteroaryl having an oxidizedring nitrogen atom is N-oxidopyridyl.

As detailed above, in some embodiments, two independent occurrences ofR⁺ (or any other variable similarly defined in the specification andclaims herein), are taken together with their intervening atom(s) toform a monocyclic or bicyclic ring selected from 3-13-memberedcycloaliphatic, 3-12-membered heterocyclyl having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, 6-10-memberedaryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

Exemplary rings that are formed when two independent occurrences of R⁺(or any other variable similarly defined in the specification and claimsherein), are taken together with their intervening atom(s) include, butare not limited to the following: a) two independent occurrences of R⁺(or any other variable similarly defined in the specification or claimsherein) that are bound to the same atom and are taken together with thatatom to form a ring, for example, N(R⁺)₂, where both occurrences of R⁺are taken together with the nitrogen atom to form a piperidin-1-yl,piperazin-1-yl, or morpholin-4-yl group; and b) two independentoccurrences of R⁺ (or any other variable similarly defined in thespecification or claims herein) that are bound to different atoms andare taken together with both of those atoms to form a ring, for examplewhere a phenyl group is substituted with two occurrences of OR⁺

these two occurrences of R⁺ are taken together with the oxygen atoms towhich they are bound to form a fused 6-membered oxygen containing ring:

It will be appreciated that a variety of other rings (e.g., Spiro andbridged rings) can be formed when two independent occurrences of R⁺ (orany other variable similarly defined in the specification and claimsherein) are taken together with their intervening atom(s) and that theexamples detailed above are not intended to be limiting.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. Additionally, unless otherwise stated, structuresdepicted herein are also meant to include compounds that differ only inthe presence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools or probes inbiological assays.

It is to be understood that, when a disclosed compound has at least onechiral center, the present invention encompasses one enantiomer ofinhibitor free from the corresponding optical isomer, racemic mixture ofthe inhibitor and mixtures enriched in one enantiomer relative to itscorresponding optical isomer. When a mixture is enriched in oneenantiomer relative to its optical isomers, the mixture contains, forexample, an enantiomeric excess of at least 50%, 75%, 90%, 95% 99% or99.5%.

The enantiomers of the present invention may be resolved by methodsknown to those skilled in the art, for example by formation ofdiastereoisomeric salts which may be separated, for example, bycrystallization; formation of diastereoisomeric derivatives or complexeswhich may be separated, for example, by crystallization, gas-liquid orliquid chromatography; selective reaction of one enantiomer with anenantiomer-specific reagent, for example enzymatic esterification; orgas-liquid or liquid chromatography in a chiral environment, for exampleon a chiral support for example silica with a bound chiral ligand or inthe presence of a chiral solvent. Where the desired enantiomer isconverted into another chemical entity by one of the separationprocedures described above, a further step is required to liberate thedesired enantiomeric form. Alternatively, specific enantiomers may besynthesized by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one enantiomer intothe other by asymmetric transformation.

When a disclosed compound has at least two chiral centers, the presentinvention encompasses a diastereomer free of other diastereomers, a pairof diastereomers free from other diasteromeric pairs, mixtures ofdiasteromers, mixtures of diasteromeric pairs, mixtures of diasteromersin which one diastereomer is enriched relative to the otherdiastereomer(s) and mixtures of diasteromeric pairs in which onediastereomeric pair is enriched relative to the other diastereomericpair(s). When a mixture is enriched in one diastereomer ordiastereomeric pair(s) relative to the other diastereomers ordiastereomeric pair(s), the mixture is enriched with the depicted orreferenced diastereomer or diastereomeric pair(s) relative to otherdiastereomers or diastereomeric pair(s) for the compound, for example,by a molar excess of at least 50%, 75%, 90%, 95%, 99% or 99.5%.

The diastereoisomeric pairs may be separated by methods known to thoseskilled in the art, for example chromatography or crystallization andthe individual enantiomers within each pair may be separated asdescribed above. Specific procedures for chromatographically separatingdiastereomeric pairs of precursors used in the preparation of compoundsdisclosed herein are provided the examples herein.

3. Description of Exemplary Compounds:

In certain embodiments, for compounds of general formula I, R², R⁴, andR⁶ are each hydrogen.

In other embodiments, R², R⁵, and R⁶ are each hydrogen.

In still other embodiments R¹ is optionally substituted C₁₋₄aliphatic,halogen, —CN, or —OMe. In yet other embodiments, R¹ is methyl, ethyl,—CF₃, Cl, —CN, —OMe, or cyclopropyl.

In still other embodiments, R² is hydrogen or —Z—R^(2a), wherein Z is—(CH₂)₂₋₄ and R^(2a) is N(R^(2b))₂) wherein each occurrence of R^(2b) isselected from hydrogen or C₁₋₄alkyl, or two occurrences of R^(2b), takentogether with the nitrogen atom to which they are bound, form anoptionally substituted C₃₋₇-heterocyclyl ring.

In yet other embodiments, R³ is methyl or CF₃.

In still other embodiments, R⁴ is methyl, or —NR^(4a)R^(4b) whereinR^(4a) and R^(4b), taken together with the nitrogen atom to which theyare bound, form an optionally substituted 3-7-membered heterocyclylring, or wherein R^(4a) is hydrogen or C₁₋₄alkyl, and R^(4b) is anoptionally substituted C₃₋₇-heterocyclyl ring or is W—R^(4c), wherein Wis an optionally substituted C₂₋₆ alkylene chain, and R^(4c) is anoptionally substituted C₃₋₇-heterocyclyl ring.

In yet other embodiments, R⁵ is an optionally substitutedC₃₋₇₋heterocyclyl ring or is X—R^(5a), wherein X is an optionallysubstituted C₂₋₆ alkylene chain, and R^(5a) is —N(R^(5b))₂, wherein eachoccurrence of R^(5b) is independently hydrogen or C₁₋₆alkyl, or twooccurrences of R^(5b), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted 3-7-membered heterocyclylring. In other embodiments, R⁴ and R⁵ are taken together to form a ringselected from:

wherein

G₁ is —NH—, —O— or —N(CH₃)—

R⁷ is selected from hydrogen or optionally substituted C₁₋₆aliphatic,

R⁸ is selected from fluoro, optionally substituted C₁₋₆aliphatic, or—YR^(1a), wherein Y is —O—, —S—, or —NR^(1a), and each occurrence ofR^(1a) is independently hydrogen, or optionally substitutedC₁₋₆aliphatic; and

y is 0-4.

In yet other embodiments, compounds of the invention have the structureof formula I-A:

In some embodiments for compounds of formula I-A, R¹ is optionallysubstituted C₁₋₄aliphatic, halogen, —CN, or —OMe. In yet otherembodiments, R¹ is methyl, ethyl, —CF₃, Cl, —CN, —OMe, or cyclopropyl.

In still other embodiments for compounds of formula I-A, R³ is methyl orCF₃.

In yet other embodiments, R⁵ is an optionally substitutedC₃₋₇₋heterocyclyl ring or is X—R^(5a), wherein X is an optionallysubstituted C₂₋₆ alkylene chain, and R^(5a) is —N(R^(5b))₂, wherein eachoccurrence of R^(5b) is independently hydrogen or C₁₋₆alkyl, or twooccurrences, of R^(5b), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted 3-7-membered heterocyclylring.

In still other embodiments, R⁵ is an optionally substitutedpyrrolidinyl, morpholinyl, piperidinyl, or piperazinyl group, or isX—R^(5a), wherein X is a C₂₋₄alkylene chain, and R^(5a) is —N(R^(5b))₂,wherein each occurrence of R^(5b) is independently hydrogen orC₁₋₆alkyl, or two occurrences of R^(5b), taken together with thenitrogen atom to which they are bound, form an optionally substitutedpyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl group. In someembodiments, the pyrrolidinyl, morpholinyl, piperidinyl, or piperazinylgroup is optionally substituted with 1-4 occurrences of C₁₋₄alkyl orC₁₋₄haloalkyl.

In still other embodiments for compounds of formula I-A:

a) R¹ is methyl, ethyl, propyl, —CF₃, Cl, —CN, —OMe, or cyclopropyl;

b) R³ is methyl or CF₃; and

c) R⁵ is an optionally substituted pyrrolidinyl, morpholinyl,piperidinyl, or piperazinyl group, or is X—R^(5a), wherein X is aC₂₋₄alkylene chain, and R^(5a) is —N(R^(5b))₂, wherein each occurrenceof R^(5b) is independently hydrogen or C₁₋₆alkyl, or two occurrences ofR^(5b), taken together with the nitrogen atom to which they are bound,form an optionally substituted pyrrolidinyl, piperidinyl, piperazinyl,or morpholinyl group.

In still other embodiments for compounds of formula R¹ is C₁ or CF₃, andR³ is methyl.

In yet other embodiments, compounds have the structure of formula I-B:

In still other embodiments, compounds have the structure of formula I-B,wherein R¹ is optionally substituted C₁₋₄aliphatic, halogen, —CN, or—OMe. In other embodiments, R¹ is methyl, ethyl, —CF₃, Cl, —CN, —OMe, orcyclopropyl.

In yet other embodiments for compounds of formula I-B, R³ is methyl orCF₃.

In still other embodiments for compounds of formula I-B, R⁴ is—NR^(4a)R^(4b), wherein R^(4a) and R^(4b), taken together with thenitrogen atom to which they are bound, form an optionally substituted3-7-membered heterocyclyl ring, or wherein R^(4a) is hydrogen orC₁₋₄alkyl, and R^(4b) is an optionally substituted C₃₋₇₋heterocyclylring or is W—R^(4c), wherein W is an optionally substituted C₂₋₄alkylene chain, and R^(4c) is an optionally substitutedC₃₋₇-heterocyclyl ring, or —N(R^(4d))₂, wherein each occurrence ofR^(4d) is independently hydrogen or C₁₋₆alkyl, or two occurrences ofR^(4d), taken together with the nitrogen atom to which they are bound,form an optionally substituted 3-7-membered heterocyclyl ring.

In other embodiments, R⁴ is —NR^(4a)R^(4b), wherein R^(4a) and R^(4b),taken together with the nitrogen atom to which they are bound, form anoptionally substituted pyrrolidinyl, piperazinyl, piperidinyl, ormorpholinyl ring, or wherein R^(4a) is hydrogen or C₁₋₄alkyl, and R^(4b)is an optionally substituted pyrrolidinyl, piperazinyl, piperidinyl, ormorpholinyl ring, or is W—R^(4c), wherein W is an optionally substitutedC₂₋₄ alkylene chain, and R^(4c) is an optionally substitutedpyrrolidinyl, piperazinyl, piperidinyl, or morpholinyl ring, or—N(R^(4d))₂, wherein each occurrence of R^(4d) is independently hydrogenor C₁₋₆alkyl, or two occurrences of R^(4d), taken together with thenitrogen atom to which they are bound, form an optionally substitutedpyrrolidinyl, piperazinyl, piperidinyl, or morpholinyl ring.

In still other embodiments for compounds of formula I-B:

a) R¹ is methyl, ethyl, propyl, —CF₃, Cl, —CN, —OMe, or cyclopropyl;

b) R³ is methyl or CF₃; and

c) R⁴ is NR^(4a)R^(4b), wherein R^(4a) and R^(4b), taken together withthe nitrogen atom to which they are bound, form an optionallysubstituted pyrrolidinyl, piperazinyl, piperidinyl, or morpholinyl ring,or wherein R^(4a) is hydrogen or C₁₋₄alkyl, and R^(4b) is an optionallysubstituted pyrrolidinyl, piperazinyl, piperidinyl, or morpholinyl ring,or is W—R^(4c), wherein W is an optionally substituted C₂₋₄ alkylenechain, and R^(4c) is an optionally substituted pyrrolidinyl,piperazinyl, piperidinyl, or morpholinyl ring, or —N(R^(4d))₂, whereineach occurrence of R^(4d) is independently hydrogen or C₁₋₆alkyl, or twooccurrences of R^(4d), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted pyrrolidinyl,piperazinyl, piperidinyl, or morpholinyl ring.

In yet other embodiments for compounds of formula I-B, R¹ is C₁ or CF₃,and R³ is methyl.

Table 1 below depicts certain exemplary compounds of formula I:

TABLE 1 Exemplary Compounds of formula I:

TABLE 2 Compound Names: I-12-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-methoxy-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-22-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-39-chloro-2-({2-methyl-6-[methyl(1-methylpyrrolidin-3-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-42-[(2,6-dimethylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-52-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-ethyl-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-69-chloro-2-({2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-79-chloro-2-({6-[3-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-89-chloro-2-({5-[3-(dimethylamino)propyl]-6-methoxypyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-92-({6-[3-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-109-chloro-2-[(5-{2-[(2R,6R)-2,6-dimethylmorpholin-4-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-119-chloro-2-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-122-({5-[4-(dimethylamino)butyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-139-chloro-2-[(6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-149-chloro-2-{[2-methyl-5-(1-methylpiperidin-4-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-159-chloro-2-[(5-{2-[3-(dimethylamino)pyrrolidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-169-chloro-2-({6-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-179-chloro-2-{[5-(1-ethylpiperidin-4-yl)-2-methylpyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-182-{[2-methyl-6-(tetrahydro-2H-pyran-4-ylamino)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-199-chloro-2-{[2-methyl-6-(tetrahydro-2H-pyran-4-ylamino)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-216-methyl-N-(2-morpholin-4-ylethyl)-5-{[6-thioxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridine-2-carboxamide I-229-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-232-[(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-249-chloro-2-({5-[3-(dimethylamino)propyl]-2-methoxypyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-252-({5-[2-(dimethylamino)ethyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-262-({2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-279-chloro-2-({5-[3-(dimethylamino)propyl]-2-fluoropyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-289-chloro-2-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-292-({6-[4-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-309-chloro-2-({5-[2-(cyclopentylamino)ethyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-319-chloro-2-[(5-{2-[(2R,6S)-2,6-dimethylmorpholin-4-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-322-({5-[3-(dimethylamino)propyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-339-chloro-10-[3-(dimethylamino)propyl]-2-(pyridin-3-ylamino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-349-chloro-2-[(5-{2-[3-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-352-({6-[[2-(dimethylamino)ethyl](methyl)amino]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-362-{[5-[3-(dimethylamino)propyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-379-chloro-2-[(2,4-dimethylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-389-chloro-2-({5-[3-(dimethylamino)propyl]-2-ethylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-399-cyclopropyl-2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-409-chloro-2-[(2-methyl-6-{[(1-methylpiperidin-4-yl)methyl]amino}pyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-419-chloro-2-({6-[[3-(dimethylamino)propyl](methyl)amino]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-429-chloro-2-({5-[3-(dimethylamino)propyl]-2,6-dimethylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-432-{[2-methyl-5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-449-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-459-chloro-2-{[2-methyl-5-(2-pyrrolidin-1-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-462-{[2-methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-479-chloro-2-[(6-pyrrolidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-499-chloro-2-({5-[(3R,5S)-4-ethyl-3,5-dimethylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-502-({5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-519-chloro-2-({5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-522-[(5-{2-[4-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-532-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-549-chloro-2-{[2-methyl-5-(2-piperidin-1-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-552-{[5-(3-hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-569-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)oxy]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-572-{[6-(isopropylamino)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-589-chloro-2-({5-[(3R)-4-ethyl-3-methylpiperazin-1-yl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-599-chloro-2-[(5-{2-[4-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-602-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-methyl-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-612-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-622-({2-methyl-5-[3-(methylamino)propyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-639-chloro-2-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-642-[(6-{[2-(dimethylamino)ethyl]amino}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-652-[(2-methyl-6-{[(1-methylpiperidin-4-yl)methyl]amino}pyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-669-chloro-2-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-672-{[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-682-[(6-aminopyridin-3-yl)amino]-9-chloro-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-699-chloro-2-({6-[4-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-709-chloro-2-{[5-[2-(dimethylamino)ethyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-719-chloro-2-({6-[[3-(dimethylamino)propyl](methyl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-729-chloro-2-{[2-methyl-5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-739-chloro-2-{[2-methyl-6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-742-({4-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-752-({2-methyl-6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-769-chloro-2-[(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-772-({2-methyl-6-[(1-methylpiperidin-4-yl)oxy]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-789-chloro-2-{[5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-792-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-6-thioxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepine-9-carbonitrile I-809-cyclopropyl-2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-819-chloro-2-[(2,6-dimethylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-829-chloro-2-[(2-methyl-6-piperidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-839-chloro-10-[3-(dimethylamino)propyl]-2-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-849-chloro-2-({5-[1-(2-fluoroethyl)piperidin-4-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-859-chloro-2-{[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-862-[(2,6-dimethyl-5,6,7,8-tetrahydro-1,6-naphthyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-872-{[2-methyl-5-(2-piperidin-1-ylethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-882-({6-[[3-(dimethylamino)propyl](methyl)amino]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-899-chloro-2-({5-[4-(dimethylamino)butyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-902-{[5-(3-aminopropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-919-chloro-2-({5-[3-(dimethylamino)propyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-922-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-939-chloro-10-[3-(dimethylamino)propyl]-2-[(2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-949-chloro-2-({5-[3-(dimethylamino)propyl]-6-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-952-({2-methyl-6-[methyl(1-methylpyrrolidin-3-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-969-chloro-2-{[6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-979-chloro-2-[(6-{[2-(dimethylamino)ethyl]amino}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-982-[(2,4-dimethylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-999-chloro-2-({5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1002-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1012-({6-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-102N-{5-[(9-chloro-6-thioxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl)amino]pyridin-2-yl}-2,2-dimethylpropanamide I-1039-chloro-2-{[2-methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1049-chloro-2-({5-[2-(dimethylamino)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1059-chloro-2-({2-methyl-6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1069-chloro-2-[(2-methyl-6-pyrrolidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1079-chloro-2-{[5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1089-chloro-2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1099-chloro-2-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1102-[(2-methyl-6-pyrrolidin-1-ylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1112-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1129-chloro-2-({6-[[2-(dimethylamino)ethyl](methyl)amino]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1132-{[(6R)-6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-1142-{[(6S)-6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneI-1152-[(2-methyl-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1165-[(9-chloro-6-thioxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl)amino]-6-methyl-N-(1-methylpiperidin-4-yl)pyridine-2-carboxamide I-1172-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-10-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-118N-[2-(dimethylamino)ethyl]-6-methyl-5-{[6-thioxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridine-2-carboxamide I-1196-methyl-N-(2-morpholin-4-ylethyl)-5-{[6-thioxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridine-2-carboxamide I-1209-chloro-10-[3-(dimethylamino)propyl]-2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1212-(dimethylamino)-N-(6-methyl-5-{[6-thioxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridin-2-yl)acetamide I-1222-{[7-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione I-1236-methyl-N-(1-methylpiperidin-4-yl)-5-{[6-thioxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridine-2-carboxamide

4. General Synthetic Methods and Intermediates:

The compounds of the present invention can be prepared by methods knownto one of ordinary skill in the art and/or by reference to the schemesshown below and the synthetic examples that follow. Exemplary syntheticroutes are set forth in Schemes 1 and 2 below, and in the Examples.

Scheme 1 above shows a general route for preparing compounds of formulaI. Conversion of anilines i to acylated amino benzoic acid methyl estersof formula ii can be accomplished by coupling with appropriatelysubstituted acyl chlorides with a suitable base, such as DIEA in thepresence or absence of DMAP. Compounds iii can be prepared from ii bycyclization with a suitable base, such as a metal alkoxide (e.g., KOt-Buor LiOt-Bu). Decarboxylation of iii to provide iv can be effected byheating in DMSO/H₂O or NMP/H₂O. For compounds iv when R¹ is an iodosubstituent, transformation of the iodide to a variety of functionalgroups can take place at this stage of the synthesis.

Treatment of compounds iv with DMF-DMA in refluxing THF is a method ofpreparing compounds v (formula II). Enamines v can be converted to thepyrimidines vi (formula IV) by treatment with appropriately substitutedpyridyl guanidines of the formula III in the presence of a mild base inethanol. Two methods for conversion of benzolactams vi to thiolactams Iare treatement with P₂S₅ in pyridine or with Lawesson's reagent.

As an alternative procedure to using pyridyl guanidines to providecompounds of formula vi, guanidine itself can be reacted with enamines vto give compounds vii (Scheme 2). Treatment of amines vii with pyridylhalides under Buchwald conditions is a method of preparing compounds offormula I.

It will be appreciated that intermediates of formulas vi, vii, and v arealso provided. In particular, intermediates of formula vi are provided,wherein variables R¹, R², R³, R⁴, R⁵ and R⁶ are as described generallyand in subsets herein as for compounds of formula I.

In one aspect, the invention provides a process for preparing a compoundof formula I:

or a salt thereof, wherein

R¹ is selected from hydrogen, —CN, halogen, optionally substitutedC₁₋₆aliphatic, or —YR^(1a),

-   -   wherein Y is —O—, —S—, or —NR^(1a), and each occurrence of        R^(1a) is independently hydrogen, or optionally substituted        C₁₋₆aliphatic;

R² is selected from hydrogen, halogen, —ZR^(2a), or —OR^(2b),

-   -   wherein Z is an optionally substituted C₁₋₆ alkylene chain, and        R^(2a) is —OR^(2b), —N(R^(2b))₂, SR^(2b), —C(O)N(R^(2b))₂,        —N(R^(2b))C(O)R^(2b), —SO₂N(R^(2b))₂, —NR^(2b)SO₂R^(2b),        —NR^(2b)C(O)N(R^(2b))₂, or NR^(2b)SO₂N(R^(2b))₂, wherein each        occurrence of R^(2b) is independently hydrogen or optionally        substituted C₁₋₆alkyl, or two occurrences of R^(2b), taken        together with a nitrogen atom to which they are bound, form an        optionally substituted 3-7-membered heterocyclyl ring;

R³ is selected from hydrogen, halogen, optionally substituted C₁₋₄alkyl,or optionally substituted C₁₋₄alkoxy;

R⁴ is selected from hydrogen, optionally substituted C₁₋₆aliphatic, anoptionally substituted 3-7-membered heterocyclyl ring,—(CH₂)_(x)NR^(4a)R^(4b), —(CH₂)_(x)NR^(4a)C(O)R^(4b),—(CH₂)_(x)NR^(4a)S(O)₂R^(4b), —(CH₂)_(x)C(O)R^(4b),—(CH₂)_(x)C(O)NR^(4a)R^(4b), —(CH₂)_(x)S(O)₂NR^(4a)R^(4b), or—(CH₂)_(x)OR^(4b),

wherein

each occurrence of x is independently 0-6;

wherein R^(4a) is hydrogen or optionally substituted C₁₋₆aliphatic, and

R^(4b) is hydrogen, optionally substituted C₁₋₆aliphatic, optionallysubstituted C₃₋₇₋heterocyclyl or C₃₋₇carbocyclyl ring, or is W—R^(4c),wherein W is an optionally substituted C₂₋₆ alkylene chain, and R^(4c)is an optionally substituted C₃₋₇-heterocyclyl ring, —OR^(4d),—N(R^(4d))₂, —SR^(4d), —C(O)N(R^(4d))₂, —N(R^(4d))C(O)R^(4d),—SO₂N(R^(4d))₂, —NR^(4d)SO₂R^(4d), —NR^(4d)C(O)N(R^(4d))₂, or—NR^(4d)SO₂N(R^(4d))₂, wherein each occurrence of R^(4d) isindependently hydrogen or optionally substituted C₁₋₆aliphatic, or twooccurrences of R^(4d), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted 3-7-membered heterocyclylring;

or wherein R^(4a) and R^(4b), taken together with the nitrogen atom towhich they are bound, form an optionally substituted 3-7-memberedheterocyclyl ring;

R⁵ is hydrogen, optionally substituted C₁₋₆aliphatic, an optionallysubstituted C₃₋₇₋heterocyclyl ring, or is X—R^(5a), wherein X is anoptionally substituted C₂₋₆ alkylene chain or —NR^(5c),

-   -   wherein when X is an optionally substituted C₂₋₆ alkylene chain        R^(5a) is —OR^(5b), —N(R^(5b))₂, —SR^(5b), —C(O)N(R^(5b))₂,        —N(R^(5b))C(O)R^(5b), —SO₂N(R^(5b))₂, —NR^(5b)SO₂R^(5b),        —NR^(5b)C(O)N(R^(5b))₂, or —NR^(5b)SO₂N(R^(5b))₂; and    -   when X is —NR^(5c), R^(5a) is hydrogen or optionally substituted        C₁₋₆aliphatic, or R^(5a) and R^(5c), taken together with the        nitrogen atom to which they are bound, form an optionally        substituted 3-7-membered heterocyclyl ring;    -   wherein each occurrence of R^(5b) and R^(5e) is independently        hydrogen or optionally substituted C₁₋₆aliphatic, or two        occurrences of R^(5b), or R^(5a) and R^(5c), taken together with        the nitrogen atom to which they are bound, form an optionally        substituted 3-7-membered heterocyclyl ring; or

wherein R⁴ and R⁵, taken together, form an optionally substituted5-7-membered cycloaliphatic or heterocyclyl ring; and

-   -   R⁶ is selected from hydrogen, halogen, optionally substituted        C₁₋₄alkyl, or optionally substituted C₁₋₄alkoxy.

The process comprises the steps of:

-   -   (a-1) treating a compound of formula II:

wherein variables R¹ and R² are as described generally and in subsetsherein as for compounds of formula I; with a compound of formula III:

wherein variables R³, R⁴, R⁵ and R⁶ are as described generally and insubsets herein as for compounds of formula I; to form a compound offormula IV:

wherein variables R¹, R², R³, R⁴, R⁵ and R⁶ are as described generallyand in subsets herein as for compounds of formula I; and

-   -   (a-2) thionating the compound of formula IV to form the compound        of formula I.

In some embodiments, the thionation of a compound of formula IV iseffected in the presence of P₂S₅ in pyridine or with Lawesson's reagent.

In another aspect, the invention provides a process for preparing acompound of formula IA:

or a salt thereof, wherein

R¹ is optionally substituted C₁₋₄aliphatic, halogen, —CN, or —OMe;

R³ is methyl or CF₃; and

R⁵ is an optionally substituted C₃₋₇₋heterocyclyl ring or is X—R^(5a),wherein X is an optionally substituted C₂₋₆ alkylene chain, and R^(5a)is —N(R^(5b)))₂, wherein each occurrence of R^(5b) is independentlyhydrogen or C₁₋₆alkyl, or two occurrences of R^(5b), taken together withthe nitrogen atom to which they are bound, form an optionallysubstituted 3-7-membered heterocyclyl ring.

The process comprises the steps of:

-   -   (b-1) treating a compound of formula II wherein R¹ is as        described generally and in subsets herein as for compounds of        formula I-A and R² is H; with an compound of formula III,        wherein variables R³ and R⁵ are as described generally and in        subsets herein as for compounds of formula I-A and R⁴ and R⁶ are        each H; to form a compound of formula IV, wherein variables R¹,        R³, and R⁵ are as described generally and in subsets herein as        for compounds of formula I-A, and R², R⁴ and R⁶ are each H; and    -   (b-2) thionating the compound of formula IV to form the compound        of formula I-A.

In a further embodiment, the process for preparing a compound of formulaI-A comprises the steps of:

-   -   (c-1) treating a compound of formula II, wherein R¹ is —CF₃ and        R² is H; with a compound of formula III, wherein R⁴ and R⁶ are        each H, R³ is methyl, R⁵ is X—R^(5a), wherein X is an optionally        substituted C₃ alkylene chain, and R^(5a) is —N(Me)₂, in the        presence of a mild base such as K₂CO₃ in a suitable solvent such        as ethanol to form a compound of formula IV; and    -   (c-2) thionating the compound of formula IV, in the presence of        P₂S₅ in pyridine to form the compound of formula I-A.

In another embodiment, the process further comprises:

-   -   (c-3) treating the compound of formula I-A with HCl in a        suitable solvent such as ethanol to form the HCl salt of formula        I-A.

In another aspect, the invention provides a process for preparing acompound of formula I-B:

or a salt thereof, wherein

R¹ is optionally substituted C₁₋₄aliphatic, halogen, —CN, or —OMe;

R³ is methyl or CF₃; and

R⁴ is —NR^(4a)R^(4b), wherein R^(4a) and R^(4b), taken together with thenitrogen atom to which they are bound, form an optionally substituted3-7-membered heterocyclyl ring, or wherein R^(4a) is hydrogen orC₁₋₄alkyl, and R^(4b) is an optionally substituted C₃₋₇₋heterocyclylring or is W—R^(4c), wherein W is an optionally substituted C₂₋₄alkylene chain, and R^(4c) is an optionally substitutedC₃₋₇-heterocyclyl ring, or —N(R^(4d))₂, wherein each occurrence ofR^(4d) is independently hydrogen or C₁₋₆alkyl, or two occurrences ofR^(4d), taken together with the nitrogen atom to which they are bound,form an optionally substituted 3-7-membered heterocyclyl ring.

The process comprises the steps of:

-   -   (d-1) treating a compound of formula II wherein R¹ is as        described generally and in subsets herein as for compounds of        formula I-B and R² is H; with an compound of formula III,        wherein variables R³ and R⁴ are as described generally and in        subsets herein as for compounds of formula I-B and R⁵ and R⁶ are        each H; to form a compound of formula IV, wherein variables R¹,        R³, and R⁴ are as described generally and in subsets herein as        for compounds of formula I-B, and R², R⁵ and R⁶ are each H; and    -   (d-2) thionating the compound of formula IV to form the compound        of formula I-B.

In yet another aspect, the invention provides a process for preparing acompound of formula III:

wherein R⁴ and R⁶ are each H, R³ is methyl, R⁵ is X—R^(5a), wherein X isan optionally substituted C₃ alkylene chain, and R^(5a) is —N(Me)₂.

The process comprises the steps of:

-   -   (e-1) treating a compound of formula V:

-   -    with a compound of formula VI:

-   -    in the presence of Pd(PPh₃)₂Cl₂, CuI, and triethylamine to give        a compound of formula VII:

-   -   (e-2) hydrogenating the compound of formula VII with hydrogen        and Pd(OH)₂ in the presence of a suitable solvent, such as        isopropyl alcohol, followed by treatment with HCl to give a        compound of formula VIII:

-   -   (e-3) treating the compound of formula VIII with a compound of        formula IX:

-   -    in the presence of acetic acid and acetonitrile to give the        compound of formula III.

Another aspect of this invention relates to compounds of formula III:

or a salt thereof, wherein variables R³, R⁴, R⁵ and R⁶ are as describedgenerally and in subsets herein as for compounds of formula I,

provided that:

i) at least one of R³, R⁴, R⁵ and R⁶ is other than H;

ii) when R³, R⁵ and R⁶ are H, then R⁴ is other than morpholinyl,—C(O)NH₂, —OMe, or —O-n-butyl;

iii) when R⁴, R⁵ and R⁶ are H, then R³ is other than chloro or —OMe; and

iv) the compound of formula III is other than:

or a salt thereof.

Another aspect of this invention relates to compounds of formula IV:

wherein variables R¹, R², R³, R⁴, R⁵ and R⁶ are as described generallyand in subsets herein as for compounds of formula I.

5. Uses, Formulation and Administration

As discussed above, the present invention provides compounds that areuseful as inhibitors of PLK enzymes, and thus the present compounds areuseful for treating proliferative, inflammatory, or cardiovasculardisorders such as tumor and/or cancerous cell growth mediated by PLK. Inparticular, the compounds are useful in the treatment of cancers in asubject, including, but not limited to, lung and bronchus, prostate,breast, pancreas, colon and recum, thyroid, liver and intrahepatic bileduct, hepatocellular, gastric, glioma/glioblastoma, endometrial,melanoma, kidney, and renal pelvis, urinary bladder, utering corpus,uterine cervix, ovary, multiple myeloma, esophagus, acute myelogenousleukemia, chronic myelogenous leukemia, lymphocytic leukemia, myeloidleukemia, brain, oral cavity, and pharynx, small intestine, non-Hodgkinlymphoma, and villous colon adenoma.

In some embodiments, compounds of the invention are suitable for thetreatment of breast cancer, bladder cancer, colon cancer, glioma,glioblastoma, lung cancer, hepatocellular cancer, gastric cancer,melanoma, thyroid cancer, endometrial cancer, renal cancer, cervicalcancer, pancreatic cancer, esophageal cancer, prostate cancer, braincancer, or ovarian cancer.

In other embodiments, compounds of the invention are suitable for thetreatment of inflammatory and cardiovascular disorders including, butnot limited to, allergies/anaphylaxis, acute and chronic inflammation,rheumatoid arthritis; autoimmunity disorders, thrombosis, hypertension,cardiac hypertrophy, and heart failure.

Accordingly, in another aspect of the present invention, pharmaceuticalcompositions are provided, wherein these compositions comprise any ofthe compounds as described herein, and optionally comprise apharmaceutically acceptable carrier, adjuvant or vehicle. In certainembodiments, these compositions optionally further comprise one or moreadditional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable prodrugs, salts,esters, salts of such esters, or any other adduct or derivative whichupon administration to a patient in need is capable of providing,directly or indirectly, a compound as otherwise described herein, or ametabolite or residue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. As used herein, the term “inhibitorily activemetabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of PLK.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺ (C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtragacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; saffloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

In yet another aspect, a method for treating a proliferative,inflammatory, or cardiovascular disorder is provided comprisingadministering an effective amount of a compound, or a pharmaceuticalcomposition to a subject in need thereof. In certain embodiments of thepresent invention an “effective amount” of the compound orpharmaceutical composition is that amount effective for treating aproliferative, inflammatory, or cardiovascular disorder, or is thatamount effective for treating cancer. In other embodiments, an“effective amount” of a compound is an amount which inhibits binding ofPLK and thereby blocks the resulting signaling cascades that lead to theabnormal activity of growth factors, receptor tyrosine kinases, proteinserine/threonine kinases, G protein coupled receptors and phospholipidkinases and phosphatases.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating the disease. The exact amountrequired will vary from subject to subject, depending on the species,age, and general condition of the subject, the severity of theinfection, the particular agent, its mode of administration, and thelike. The compounds of the invention are preferably formulated in dosageunit form for ease of administration and uniformity of dosage. Theexpression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disease beingtreated and the severity of the disease; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

While one or more of the inventive compounds may be used in anapplication of monotherapy to treat a disorder, disease or symptom, theyalso may be used in combination therapy, in which the use of aninventive compound or composition (therapeutic agent) is combined withthe use of one or more other therapeutic agents for treating the sameand/or other types of disorders, symptoms and diseases. Combinationtherapy includes administration of the therapeutic agents concurrentlyor sequentially. Alternatively, the therapeutic agents can be combinedinto one composition which is administered to the patient.

In one embodiment, the compounds of this invention are used incombination with other therapeutic agents, such as other inhibitors ofPLK. In some embodiments, a compound of the invention is administered inconjunction with a therapeutic agent selected from the group consistingof cytotoxic agents, radiotherapy, and immunotherapy. It is understoodthat other combinations may be undertaken while remaining within thescope of the invention.

Another aspect of the invention relates to inhibiting PLK, activity in abiological sample or a patient, which method comprises administering tothe patient, or contacting said biological sample with a compound offormula I or a composition comprising said compound. The term“biological sample”, as used herein, generally includes in vivo, invitro, and ex vivo materials, and also includes, without limitation,cell cultures or extracts thereof; biopsied material obtained from amammal or extracts thereof; and blood, saliva, urine, feces, semen,tears, or other body fluids or extracts thereof.

Still another aspect of this invention is to provide a kit comprisingseparate containers in a single package, wherein the inventivepharmaceutical compounds, compositions and/or salts thereof are used incombination with pharmaceutically acceptable carriers to treatdisorders, symptoms and diseases where PLK kinase plays a role.

Experimental Procedures Definitions AcOH acetic acid AHX aminohexanoicacid ATP adenosine triphosphate Bn benzyl BOC tert-butoxycarbonyl BSAbovine serum albumin C. Celsius CO₂ Carbon Dioxide DBU1,8-diazabicyclo-undec-7-ene DCM dichloromethane DIEAdiisopropylethylamine DMAP 4-dimethylaminopyridine DMF dimethylformamideDMF-DMA dimethylformamide dimethylacetal DMSO dimethylsulfoxide DNAdeoxyribonucleic acid DTT dithiothreitol EDCN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride EDTAethylenediaminetetraacetic acid EtOAc ethyl acetate EtOH ethanol FBSfetal bovine serum h hours IC₅₀ inhibitory concentration 50% IgGimmunoglobulin G KCl Potassium Chloride L length Lawesson's2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,3,2,4-dithiadi- Reagentphosphetane m/z mass to charge MeOH methanol min minutes MS massspectrum NaCl Sodium Chloride Ni Nickel NMP N-methylpyrrolidinone PBSphosphate-buffered saline PBST phosphate-buffered saline Tween20 pHisH3phosphorylated histone H3 on Serine 10 PLK1 polo-like kinase 1 rt roomtemperature t-BuOK potassium tert-butoxide STAB sodium triacetoxyborohydride TBDPS tert-butyl diphenyl silyl TBS tert-butyl dimethylsilyl TEA triethylamine THF tetrahydrofuran TBTUO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate Wwidth Xphos 2-dicyclohexylphosphino-2′,4′,6′-tri-i-propyl-1,1′-biphenyl

LCMS Conditions:

Method Formic Acid (FA): Spectra were run on a Phenominex Luna 5μ C1850×4.6 mm column on a Hewlett-Packard HP 1100 using acetonitrilecontaining zero to 100 percent 0.1% formic acid in water (2.5 mL/min fora 3 min run).

Method Formic Acid Long (FAL): Spectra were run on a Phenominex Luna 5μC18 50×4.6 mm column on a Hewlett-Packard HP1100 using acetonitrilecontaining zero to 100 percent 0.1% formic acid in water (1.0 mL/min fora 16 min run).

Method Ammonium Acetate (AA): Spectra were run on a Phenominex Luna 5μC18 50×4.6 mm column on a Hewlett-Packard HP1100 using acetonitrilecontaining zero to 100 percent 10 mM ammonium acetate in water (2.5mL/min for a 3 min run).

Method Ammonium Acetate Long (AAL): Spectra were run on a PhenominexLuna 5μ C18 50×4.6 mm column on a Hewlett-Packard HP1100 usingacetonitrile containing zero to 100 percent 10 mM ammonium acetate inwater (1.0 mL/min for a 16 min run).

For the LCMS Methods FAP1, FAP2, and FAP3, Solvent A=99% Water+1%Acetonitrile+0.1% Formic Acid and Solvent B=95% Acetonitrile+5%Water+0.1% Formic Acid.

Method Formic Acid Purity 1 (FAP1): Spectra were obtained on an Agilent1100 Series LC system connected to a Micromass mass spectrometer usingWaters Symmetry C-18 4.6×100 mm column with a solvent gradient of 100% Ato 100% B (1 mL/min for a 20 min run).

Method Formic Acid Purity 2 (FAP2): Spectra were obtained on an Agilent1100 Series LC system connected to a Micromass mass spectrometer usingWaters Symmetry C-18 4.6×100 mm column with a solvent gradient of 100% Ato 100% B (1 mL/min for a 16.5 min run).

Method Formic Acid Purity 3 (FAP3): Spectra were obtained on Agilent1100 Series LC system connected to a Micromass mass spectrometer usingWaters Symmetry C-18 4.6×100 mm column with a solvent gradient of 95%A/5% B to 100% B (1 mL/min for a 10 min run).

Example 1 Synthesis of Compounds of Formula i 2-Amino-4-methoxy-benzoicacid methyl ester

Step 1: 4-Iodo-2-nitrobenzoic acid

To a mixture of 4-iodo-2-nitrotoluene (9.0 g, 34 mmol) in H₂O (340 mL)was added KMnO₄ (22.0 g, 139 mmol). The mixture was heated at reflux for5 h and then cooled to rt and filtered through Celite®. The filtrate wasacidified to pH=2 with conc. HCl. The precipitate that formed and wasfiltered and dried to give 4-iodo-2-nitrobenzoic acid (2.0 g, 20%). Thefiltrate was extracted with DCM (3×200 mL). The organic solutions werecombined, dried over Na₂SO₄, filtered, and concentrated to provide asecond lot of 4-iodo-2-nitrobenzoic acid (0.18 g, 2%).

Step 2: Methyl 4-iodo-2-nitrobenzoate

To a solution of 4-iodo-2-nitrobenzoic acid (2.3 g, 7.9 mmol) in DMF (30mL) was added DBU (2.4 mL, 16 mmol) followed by iodomethane (1.5 mL, 24mmol). The reaction mixture was allowed to stir at 0° C. for 15 min,then slowly allowed to warm to rt and to stir overnight. After 12 h, thereaction mixture was poured into 200 mL of H₂O and extracted with EtOAc(2×100 mL). The organic solutions were combined, washed with H₂O (2×100mL), dried over Na₂SO₄, filtered and concentrated to give methyl4-iodo-2-nitrobenzoate (2.3 g, 96%).

Step 3: Methyl 2-amino-4-iodobenzoate

To a solution of methyl 4-iodo-2-nitrobenzoate (2.3 g, 7.4 mmol) in DCM(5 mL) and EtOAc (5 mL) was added SnCl₂-2H₂O (10.8 g, 89.7 mmol). Themixture was allowed to stir for 12 h at rt. The solvents wereevaporated, and the residue was partitioned between 200 mL of saturatedaqueous NaHCO₃ and 200 mL of DCM. The organic solution was separated andthe aqueous solution was extracted with DCM (2×100 mL). The organicsolutions were combined, dried over Na₂SO₄, filtered and concentrated togive methyl 2-amino-4-iodobenzoate (1.85 g, 90%).

Methyl 2-amino-4-methoxybenzoate Step 1: Methyl4-methoxy-2-nitrobenzoate

To a solution of 4-methoxy-2-nitrobenzoic acid (10.0 g, 50.7 mmol) inDMF (200 mL) at 0° C. was added DBU (15.2 mL, 101 mmol) followed byiodomethane (9.47 mL, 152 mmol). The reaction mixture was allowed tostir at 0° C. for 15 min then at rt overnight. The mixture was pouredinto water and extracted with EtOAc. The organic solutions werecombined, washed with brine and dried over MgSO₄. The residue waspurified by column chromatography to give methyl4-methoxy-2-nitrobenzoate (50.7 mmol, 85%) as a yellow solid. LCMS (FA):m/z=212.1 (M+H).

Step 2: Methyl 2-amino-4-methoxybenzoate

A solution of methyl 4-methoxy-2-nitrobenzoate (9.0 g, 43 mmol) in MeOH(100 mL) was degassed and purged with nitrogen. To this solution wasadded 10% palladium hydroxide on carbon (1.2 g). The reaction mixturewas degassed, purged with hydrogen and allowed to stir at rt overnight.The reaction mixture was filtered through Celite®, the Celite was washedwith MeOH and the filtrate was concentrated to give methyl2-amino-4-methoxybenzoate (7.80 g, 76%) as a brown solid. LCMS (FA):m/z=182.1 (M+H).

Methyl 2-amino-4-(trifluoromethyl)benzoate

Step 1: 2-Amino-4-(trifluoromethyl)benzoic acid

A mixture of 2-nitro-4-(trifluoromethyl)benzoic acid (646 g, 2.75 mol)and 10% palladium on carbon (50% in water, 71 g) in MeOH (3.6 L) wasallowed to stir at rt under an atmosphere of H₂ (15 psi). After 3 h, TLC(DCM:MeOH 10:1) showed no remaining starting material. The mixture wasfiltered and the filtrate was concentrated to give2-amino-4-(trifluoromethyl)benzoic acid (520 g, 92%) as a white solid.

Step 2: Methyl 2-amino-4-(trifluoromethyl)benzoate

To a mixture of 2-amino-4-(trifluoromethyl)benzoic acid (150 g, 0.73mol) in MeOH (2.5 L) was added conc. HCl (0.5 L, 16.5 mol). The reactionmixture was allowed to stir at reflux. After 3 h, an additional portionof HCl (0.5 L, 16.5 mol) was added and the reaction was allowed tocontinue to stir at 80° C. for 60 h. The reaction mixture was allowed tocool to rt and was concentrated. Water (0.5 L) was added to the residue,and the solution was basified with 10% aqueous NaOH solution. Theresulting precipitate was filtered to give methyl2-amino-4-(trifluoromethyl)benzoate (120 g, 75%) as a white solid.

Methyl 2-amino-4-chlorobenzoate

To a mixture of 2-amino-4-chlorobenzoic acid (150 g, 0.88 mol) in MeOH(2.6 L) was added conc. HCl (0.5 L, 16.5 mol) and the reaction mixturewas allowed to stir at reflux. After 12 h, the reaction mixture wasallowed to cool to rt and was concentrated. Water (0.5 L) was added tothe residue and the solution basified with 10% aqueous NaOH. Theresulting precipitate was filtered to give methyl2-amino-4-chlorobenzoate (101 g, 62%).

Methyl 2-amino-4-chloro-5-iodobenzoate

To a mixture of iodine (68 g, 0.27 mol) and silver sulfate (84 g, 0.27mol) in absolute EtOH (2.5 L) was added methyl 2-amino-4-chlorobenzoate(50 g, 0.27 mol). The reaction mixture was allowed to stir at rt for 45min. The reaction mixture was then filtered through a pad of Celite® andthe filtrate was concentrated. The residue was dissolved in EtOAc (2 L)and washed with saturated aqueous NaHCO₃ (3×400 mL), water (3×400 mL),and brine. The organic solution was dried over Na₂SO₄, filtered, andconcentrated to give methyl 2-amino-4-chloro-5-iodobenzoate (85 g, 99%).

Example 2 Synthesis of Compounds of Formula iv8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione

Step 1: Methyl 4-iodo-2-[(4-methoxy-4-oxobutanoyl)amino]benzoate

To a solution of methyl 2-amino-4-iodobenzoate (17 g, 61.3 mmol) in DCM(200 mL), was added DIEA (10.6 mL, 64.4 mmol) and DMAP (37.5 mg, 0.31mmol). To this solution was added 3-(carbomethoxy)propionyl chloride(8.3 mL, 67.4 mmol) dropwise, and the reaction mixture was allowed tostir at rt for 2 h. To the reaction mixture was then added H₂O (80 mL)and the mixture was allowed to stir for 30 min. The organic solution wasseparated and the aqueous solution was extracted with DCM (2×100 mL).The organic solutions were combined, washed with H₂O (2×100 mL) andbrine (1×100 mL), dried over Na₂SO₄, filtered and concentrated to givemethyl 4-iodo-2-[(4-methoxy-4-oxobutanoyl)amino]benzoate (24.6 g, 99%).

Step 2: Methyl8-iodo-2,5-dioxo-2,3,4,5-tetrahydro-1H-1-benzazepine-4-carboxylate

To a solution of methyl4-iodo-2-[(4-methoxy-4-oxobutanoyl)amino]benzoate (24.6 g, 63 mmol) inTHF (240 mL) at 10° C. was added a 1 M solution of KOt-Bu in THF (185mL, 185 mmol) dropwise over 30 min while maintaining the temperature at10° C. After 2.5 h, 50 mL of H₂O followed by 190 mL of 1N HCl were addedto bring the solution to pH=4. The resulting mixture was allowed to stirat rt for 40 min. The organic solution was separated and the aqueoussolution was extracted with EtOAc (2×200 mL). The organic solutions werecombined, dried over Na₂SO₄, filtered and concentrated to give methyl8-iodo-2,5-dioxo-2,3,4,5-tetrahydro-1H-1-benzazepine-4-carboxylate (22g, 97%).

Step 3: 8-Iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione

A mixture of methyl8-iodo-2,5-dioxo-2,3,4,5-tetrahydro-1H-1-benzazepine-4-carboxylate (74g, 0.2 mol) in DMSO (560 mL) and H₂O (16 mL) was heated at 150° C. for 4h. The reaction mixture was allowed to cool to rt, ice (1.0 L) wasadded, and the mixture was allowed to stir 12 h. To the flask was added1N HCl (1.0 L) at 0° C. and the mixture allowed to stir for 3 h. Theresulting precipitate was filtered and dried under reduced pressure toafford 8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione (60 g, 97%).

8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione

Step 1: Methyl 2-(4-methoxy-4-oxobutanamido)-4-(trifluoromethyl)benzoate

A 22 L reactor equipped with an overhead stirrer and a temperature probewas charged with methyl 2-amino-4-(trifluoromethyl)benzoate (680 g, 3.1mol). DCM (5.4 L, 8 vol) was added to it and the resulting solution wascooled in an ice-water bath. DIEA (1.08 L, 6.2 mol, 2 equiv) was addedto the solution. A solution of 4-chloro-4-oxo-butyrate (568 mL, 4.65mol, 1.5 equiv) in DCM (1.4 L, 2 vol) was added dropwise maintaining theinternal temperature below 15° C. (3 h). The ice-water bath was removedand the reaction was stirred for 3 h. The reaction was judged completeby HPLC analysis. Water (3.4 L, 5 vol) was added to the reaction and thebiphasic mixture was stirred at ambient temperature overnight. The waterlayer was removed and saturated aqueous NaHCO₃ (3.4 L, 5 vol) was addedto the DCM layer. The mixture was stirred for 30 min. The two layerswere separated and another 3.4 L (5 vol) of saturated aqueous NaHCO₃ wasadded to the DCM layer. The mixture was stirred for 30 min. The twolayers were separated and a 1:1 mixture of brine/water (3.4 L, 5 vol)was added to the DCM layer. The biphasic mixture was stirred for 30 min.The two layers were separated and the DCM layer was pumped down to a lowvolume (ca. 1.4 L, 2 vol). THF (6.8 L, 10 vol) was added to the solutionand the solvent was evaporated under reduced pressure to a low volume(ca. 1.4 L) to obtain Methyl2-(4-methoxy-4-oxobutanamido)-4-(trifluoromethyl)benzoate [1.032 kg,theoretical yield, 3.1 mol] as a solution in THF.

Step 2: Methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylate

A 22 L reactor equipped with an overhead stirrer and a temperature probewas charged with LiOBu^(t) (1.24 kg, 15.5 mol, 5 equiv), and THF (5 L, 5vol) was added slowly to obtain a solution (1 h). The internaltemperature rose to 28° C. during this time. The solution was stirreduntil the temperature subsided to 22° C. The solution of Methyl2-(4-methoxy-4-oxobutanamido)-4-(trifluoromethyl)benzoate [3.1 mol] inTHF obtained above was further diluted with THF to make the total volume5 L. This solution was added dropwise to the reactor maintaining theinternal temperature below 35° C. (addition time 2 h). The reaction wasstirred overnight at ambient temperature and was judged complete by HPLCanalysis (Method 1, 15 min method). The reactor was cooled in anice-water bath. The reaction was quenched by adding a mixture of AcOH(1.33 L, 23.25 mol, 7.5 equiv) and water (5 L, 5 vol) maintaining theinternal temperature below 35° C. The pH of resulting biphasic mixturewas 6-7. THF was removed resulting in a brown suspension. The solid wascollected by filtration and the filter cake was washed with 5% THF inwater (4 L, 4 vol). The solid was dried in a vacuum oven at 35° C.overnight to obtain Methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylate[970 g, 103% recovery] as a brown solid. HPLC purity of this solid was89.2% (AUC @ 226 nm).

Crude methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylateobtained above was purified by recrystallization from IPA. Thus, a 12 Lreactor equipped with a reflux condenser was charged with crude methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylate(970 g) and IPA (6.8 L, 7 vol). The suspension was heated at 80° C. for1.5 h resulting in a brown slurry. This was then cooled slowly toambient temperature over a period of 3 h. The suspension that resultedwas further cooled to −15° C. by using an ice-methanol bath. Thesuspension was stirred at this temperature for 2 h. The solid wasfiltered and the filter cake was washed with cold IPA (3.8 L, cooled inice-methanol bath). The solid was dried in a vacuum oven at 35° C. to aconstant weight to obtain methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylate[671 g, 72% yield over two steps) as an off-white solid. ¹H NMR (500MHz, DMSO-d₆) δ ppm: 12.37 (s, 1H), 10.55 (s, 1H), 8.01 (d, 1H, J=10Hz), 7.59 (d, 1H, J=10 Hz), 7.53 (s, 1H), 3.85 (s, 3H), 2.99 (s, 2H);ESI-MS m/z: 302 (M+H, 100%).

Step 3: 8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione

A 22 L reactor was charged with a solution of methyl2,5-dioxo-8-(trifluoromethyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-4-carboxylate[670 g, 2.22 mol] in NMP (4 L, 6 vol). Water (67 mL, 0.1 vol) was addedand the resulting solution was slowly heated to 135° C. The reaction wascontinued for 15 h when HPLC analysis indicated >99% conversion. Thereaction was cooled to ambient temperature. Water (8.7 L, 13 vol) wasadded slowly with vigorous stirring resulting in a suspension. Thesuspension was stirred at ambient temperature overnight. The solid wascollected by filtration and the filter cake was washed with water (1.5L). The solid was dried to a constant weight in a vacuum oven at 45° C.to obtain 8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione(522 g, 97% recovery) as an off-white solid. ¹H NMR (500 MHz, DMSO-d₆) δppm: 10.28 (s, 1H), 8.00 (d, 1H, J=8.0 Hz), 7.54 (s, 1H), 7.50 (d, 1H,J=8.0 Hz), 3.00-2.94 (m, 2H), 2.78-2.70 (m, 2H); ESI-MS m/z: 244 (M+H,100%).

Compounds in the following table may be prepared from the appropriatestarting materials using the procedures described above:

8-methoxy-3,4-dihydro-1H-1-benzazepine-2,5-dione8-chloro-3,4-dihydro-1H-1-benzazepine-2,5-dione8-chloro-7-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione

8-Methyl-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a degassed solution of 8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione(0.3 g, 1.0 mmol) in DMF (5 mL) was added CuI (19 mg, 0.1 mmol), KF (116mg, 2.0 mmol), Me₄Sn (0.28 mL, 2.0 mmol), and Pd(PPh₃)₄ (116 mg, 0.1mmol). The reaction mixture was allowed to stir at 120° C. for 4 h.After being allowed to cool to rt, EtOAc (50 mL) and a 1M aqueous KFsolution (25 mL) were added and the solution was allowed to stir for anadditional 40 min. The mixture was filtered over Celite®. The organicsolution was separated, washed with brine, dried over Na₂SO₄, filteredand concentrated. The residue was purified by column chromatography toprovide 8-methyl-3,4-dihydro-1H-1-benzazepine-2,5-dione (60 mg, 32%).

8-Ethyl-3,4-dihydro-1H-1-benzazepine-2,5-dione

Step 1: 8-Vinyl-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a degassed solution of 8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione(0.3 g, 1.0 mmol) in DMF (5 mL) was added CuI (19 mg, 0.1 mmol), KF (116mg, 2.0 mmol), tributylethenylstannane (0.58 mL, 2.0 mmol), andPd(PPh₃)₄ (116 mg, 0.1 mmol). The reaction mixture was allowed to stirat 100° C. for 4 h. After being allowed to cool to rt, EtOAc (50 mL) anda 1M aqueous KF solution (25 mL) were added and the mixture was allowedto stir for an additional 40 min. The mixture was filtered over Celite®and the organic solution was separated, washed with brine, dried overNa₂SO₄, filtered and concentrated. The residue was treated withn-pentane and filtered. The resulting solid was purified by columnchromatography to provide 8-vinyl-3,4-dihydro-1H-1-benzazepine-2,5-dione(80 mg, 40%).

Step 2: 8-Ethyl-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a solution of 8-vinyl-3,4-dihydro-1H-1-benzazepine-2,5-dione (2.6 g,13 mmol) in EtOH (160 mL) was added 10% (w/w) palladium on carbon (260mg). The reaction mixture was allowed to stir under an atmosphere of H₂(5 bar) at 70° C. until reaction was complete. The reaction mixture wasfiltered over Celite® and the filtrate was concentrated to give8-ethyl-3,4-dihydro-1H-1-benzazepine-2,5-dione (2.4 g, 91%).

8-Cyclopropyl-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a solution of 8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione (0.6 g,2.0 mmol) in toluene (9 mL) and H₂O (1 mL) under an atmosphere of N₂ wasadded cyclopropylboronic acid (0.34 g, 4.0 mmol), K₃PO₄ (1.27 g, 6.0mmol), PCy₃ (1.2 mL, 0.8 mmol), and Pd(OAc)₂ (90 mg, 0.4 mmol). Thereaction mixture was allowed to stir at 80° C. for 12 h and then allowedto cool to rt. To the mixture were added EtOAc (50 mL) and H₂O (50 mL),and the resulting solids were filtered over Celite®. The organicsolution was separated and the aqueous solution was extracted with EtOAc(2×50 mL). The organic solutions were combined, dried over MgSO₄,filtered and concentrated. The residue was purified by columnchromatography to provide8-cyclopropyl-3,4-dihydro-1H-1-benzazepine-2,5-dione (100 mg, 23%).

8-Cyano-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a solution of 8-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione (0.3 g,1.0 mmol) in DMF (5 mL) was added Zn(CN)₂ (0.12 g, 1.0 mmol) andPd(PPh₃)₄ (116 mg, 0.1 mmol). The reaction mixture was allowed to stirat 80° C. for 3.5 h. After being allowed to cool to rt, EtOAc (20 mL)and H₂O (20 mL) were added, the organic solution was separated, and theaqueous solution was extracted with EtOAc (2×50 mL). The organicsolutions were combined, washed with H₂O and brine, dried over MgSO₄,filtered and concentrated. The residue was purified by columnchromatography to provide 8-cyano-3,4-dihydro-1H-1-benzazepine-2,5-dione(140 mg, 70%).

Example 3 Synthesis of Compounds of Formula v (Formula II)(4Z)-8-chloro-4-[(dimethylamino)methylene]-7-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione

To a suspension of8-chloro-7-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione (10.0 g, 29.8mmol) in THF (44 mL) was added DMF-DMA (21.1 mL, 149 mmol). The flaskreaction mixture was allowed to stir 60° C. under an atmosphere ofargon. A light orange solution containing a suspended solid resulted.After 17 h, the reaction mixture was allowed to cool to rt. Ether (100mL) was added and the solid was collected via suction filtration, washedwith ether, and dried in a vacuum oven to yield(4Z)-8-chloro-4-[(dimethylamino)methylene]-7-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione(9.50 g, 24.3 mmol, 82%) as a yellow solid. LCMS (FA): m/z=391 (M+H).

(4Z)-4-[(dimethylamino)methylene]-8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione

A 250 mL, round-bottom flask equipped with a magnetic stir bar and areflux condenser was charged with8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione [5 g, 0.02mol] and THF (50 mL) was added to it. DMF-DMA (13.4 mL, 0.1 mol, 5equiv) was added to the resulting suspension; the reaction was heated at60° C. for 3 h and was judged complete by HPLC analysis. The reactionmixture was cooled to ambient temperature and the yellow solid thatformed was collected by filtration. The filter cake was washed with 1:1THF/MTBE (25 mL, 5 vol) followed by MTBE (10 mL). The solid was driedunder high vacuum to obtain enamine(4Z)-4-[(dimethylamino)methylene]-8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione(5.28 g, 88% yield) as a yellow solid. ¹H NMR (500 MHz, DMSO-d₆) δ ppm:10.14 (s, 1H), 7.90 (d, 1H, J=8.0 Hz), 7.67 (s, 1H), 7.47 (d, 1H, J=8.0Hz), 7.37 (s, 1H), 3.3 (s, 2H), 3.24 (s, 6H); ESI-MS m/z: 299 (M+H,100%).

Compounds in the following table may be prepared from the appropriatestarting materials using the procedures described above:

(4Z)-4-[(dimethylamino)methylene]-8-methoxy-3,4-dihydro-1H-1-benzazepine-2,5-dione(4Z)-8-chloro-4-[(dimethylamino)methylene]-3,4-dihydro-1H-1-benzazepine-2,5-dione(4Z)-4-[(dimethylamino)methylene]-2,5-dioxo-2,3,4,5-tetrahydro-1H-1-benzazepine-8-carbonitrile(4Z)-4-[(dimethylamino)methylene]-8-ethyl-3,4-dihydro-1H-1-benzazepine-2,5-dione(4Z)-8-cyclopropyl-4-[(dimethylamino)methylene]-3,4-dihydro-1H-1-benzazepine-2,5-dione

Example 4 Synthesis of Compounds of Formula vi (Formula IV)9-Chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To(4E)-8-chloro-4-[(dimethylamino)methylene]-3,4-dihydro-1H-1-benzazepine-2,5-dione(3.25 g. 12.3 mmol) in EtOH (50 mL) were addedN-{2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}guanidine*xHC1 (6.5 g, 14 mmol) and potassium carbonate (13.6 g, 98.3 mmol). Thereaction mixture was allowed to stir for 2 days at 70° C. and thenallowed to cool to room temperature. The solids were filtered and thefiltrate was concentrated to give a thick red oil. Water was added tothe residue and the aqueous solution was extracted with EtOAc. Theorganic solutions were combined, dried, filtered and concentrated togive9-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(5.3 g, 93%).

2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

N-{5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}guanidine.triHCl(233 g, 676 mol) was dissolved in ethanol (1.42 L) and to it was addedpowder potassium carbonate (359 g, 2.60 mol). The mixture was stirred atroom temperature for 15 min.(4Z)-4-[(dimethylamino)methylene]-8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione(155 g, 520 mmol) was added to the reaction mixture. This mixture wasslowly heated at 70° C. and held at 70° C. for 30 min. The reactionmixture was heated at 75° C. for 18 hours. HPLC indicated competition ofthe reaction. The reaction mixture was cooled to room temperature andfiltered to remove potassium carbonate. The cake was washed with ethanol(200 mL) and the filtrate was concentrated to low volume (1.1 L). Themixture was heated at 70° C. while water (2.0 L) was slowly added. Thesuspension formed was stirred at 75° C. for 1 hour, then slowly cooledto room temperature. This solid was filtered and washed with 300 mLwater/EtOH (2-vol/1 vol). Drying under vacuo at 40° C. overnightafforded2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-oneas a light pink solid (175 g, >98% pure, 71.6% yield). LCMS (FA):R_(t)=5.72 min, m/z=469.2 (M−H).

Compounds in the following table may be prepared from the appropriatestarting materials using the procedure described above:

2-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridin-3-yl]amino}-9-(trifluoro-methyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-methoxy-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-2-methoxypyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-2-fluoropyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-10-[3-(dimethylamino)propyl]-2-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-cyclopropyl-2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-ethyl-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-6-oxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepine-9-carbonitrile2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-methyl-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-cyclopropyl-2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[3-(dimethylamino)propyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(2-piperidin-1-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[2-methyl-5-(2-piperidin-1-ylethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[2-methyl-5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(5-{2-[3-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(2-pyrrolidin-1-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[4-(dimethylamino)butyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[4-(dimethylamino)butyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(5-{2-[4-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(5-{2-[4-(dimethylamino)piperidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(5-{2-[(2R,6R)-2,6-dimethylmorpholin-4-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(5-{2-[(2R,6S)-2,6-dimethylmorpholin-4-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(5-{2-[3-(dimethylamino)pyrrolidin-1-yl]ethyl}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({6-[4-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-6-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[6-(isopropylamino)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[2-methyl-6-(tetrahydro-2H-pyran-4-ylamino)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({2-methyl-6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(6-{[2-(dimethylamino)ethyl]amino}-2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(6-{[2-(dimethylamino)ethyl]amino}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({6-[[2-(dimethylamino)ethyl](methyl)amino]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[[2-(dimethylamino)ethyl](methyl)amino]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[[3-(dimethylamino)propyl](methyl)amino]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({6-[[3-(dimethylamino)propyl](methyl)amino]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-6-[methyl(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-6-(tetrahydro-2H-pyran-4-ylamino)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[3-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({6-[3-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({6-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-6-[(1-methylpiperidin-4-yl)oxy]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2,4-dimethylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(2,4-dimethylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(2,6-dimethylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2,6-dimethylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(2-methyl-6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(2-methyl-6-piperidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(2-methyl-6-piperidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2-methyl-6-pyrrolidin-1-ylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({2-methyl-6-[(1-methylpiperidin-4-yl)oxy]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[4-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-6-[methyl(1-methylpyrrolidin-3-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({2-methyl-6-[methyl(1-methylpyrrolidin-3-yl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(1-methylpiperidin-4-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[5-(1-ethylpiperidin-4-yl)-2-methylpyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[1-(2-fluoroethyl)piperidin-4-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-2-ethylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({2-methyl-5-[3-(methylamino)propyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one6-methyl-N-(2-morpholin-4-ylethyl)-5-{[6-oxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridine-2-carboxamide9-chloro-2-[(2-methyl-6-{[(1-methylpiperidin-4-yl)methyl]amino}pyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2-methyl-6-{[(1-methylpiperidin-4-yl)methyl]amino}pyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({4-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-[(2,6-dimethyl-5,6,7,8-tetrahydro-1,6-naphthyridin-3-yl)amino]-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[5-(3-hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-{[2-methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[2-methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[(3R,5S)-4-ethyl-3,5-dimethylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

9-Chloro-2-({5-[3-(dimethylamino)propyl]-6-methoxypyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A mixture of2-amino-9-chloro-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one (0.36g, 1.38 mmol),3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropan-1-amine (0.38 g,1.38 mmol), xphos (0.047 g, 0.099 mmol), andtris(dibenzylideneacetone)dipalladium(0) (0.023 g, 0.026 mmol) in asealed microwave tube was evacuated and purged with nitrogen threetimes. To the solids were added tert-butyl alcohol (3.6 mL) and t-BuOK(1M in t-BuOH, 2.63 mL) via syringe. The mixture was stirred well andthen subjected to microwave irradiation (150 watts) while heating at150° C. for 45 min. The reaction mixture was then poured into water (25mL) with vigorous stirring. The precipitate that formed was filtered,washed with water, and air dried. The residue was purified by columnchromatography to give9-chloro-2-({5-[3-(dimethylamino)propyl]-6-methoxypyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.125 g, 18%).

Compounds in the following table were prepared from the appropriatestarting materials using the procedures described above:

2-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-[(6-morpholin-4-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({6-[[3-(dimethylamino)propyl](methyl)amino]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-oneN-{5-[(9-chloro-6-oxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl)amino]pyridin-2-yl}-2,2-dimethylpropanamide9-chloro-2-[(6-pyrrolidin-1-ylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[5-[2-(dimethylamino)ethyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[2-(cyclopentylamino)ethyl]-2-methylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one2-({5-[2-(dimethylamino)ethyl]pyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[2-(dimethylamino)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[5-(2-morpholin-4-ylethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-{[5-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[(3R)-4-ethyl-3-methylpiperazin-1-yl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]pyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

9-Chloro-10-[3-(dimethylamino)propyl]-2-[(2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

Step 1:2-Amino-9-chloro-10-iodo-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A suspension of(4Z)-8-chloro-4-[(dimethylamino)methylene]-7-iodo-3,4-dihydro-1H-1-benzazepine-2,5-dione(9.50 g, 24.3 mmol), guanidine hydrochloride (2.56 g, 26.8 mmol), andpotassium carbonate (11.1 g, 80.3 mmol) in EtOH (143) was allowed tostir while heating at 70° C. in a sealed reaction vessel. After 19 h,the reaction mixture was allowed to cool to room temperature then addedto water (200 mL) and allowed to stir for 1 h. The resulting tan solidwas collected via suction filtration, washed with water, and dried in avacuum oven to give2-amino-9-chloro-10-iodo-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(7.69 g, 82%). LCMS (FA): m/z=387 (M+H).

Step 2:2-Amino-9-chloro-10-[3-(dimethylamino)prop-1-yn-1-yl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A solution of2-amino-9-chloro-10-iodo-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(1.66 g, 4.29 mmol) and propargyl(dimethylamine) (0.74 mL, 6.89 mmol) intriethylamine (7 mL) and DMF (7 mL) was degassed with argon. To thissolution was added bis(triphenylphosphine)palladium(II) chloride (0.30g, 0.43 mmol) and copper iodide (0.163 g, 0.86 mmol). The reactionmixture was allowed to stir at 75° C. overnight. After the reactionmixture was allowed to cool to rt, water was added. The precipitate thatformed was filtered and dried to give2-amino-9-chloro-10-[3-(dimethylamino)prop-1-yn-1-yl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(1.61 g, 98%) as a brown solid. LCMS (FA): m/z=342 (M+H).

Step 3:2-Amino-9-chloro-10-[3-(dimethylamino)propyl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A slurry of Raney Ni (3.0 mL) in water was added to2-amino-9-chloro-10-[3-(dimethylamino)prop-1-yn-1-yl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.30 g, 0.88 mmol) in THF (6 mL). The reaction mixture was allowed tostir under an atmosphere of H₂ at rt overnight. The reaction mixture wasfiltered over Celite and the filter cake was washed with THF. Thefiltrate was concentrated and the resulting solid was triturated withMeOH, diethyl ether, and hexanes. The solid was collected via suctionfiltration to give2-amino-9-chloro-10-[3-(dimethylamino)propyl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.158 g, 52%) as a light yellow solid. LCMS (FA): m/z=346 (M+H).

Step 4:9-Chloro-10-[3-(dimethylamino)propyl]-2-[(2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To a mixture of2-amino-9-chloro-10-[3-(dimethylamino)propyl]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.10 g, 0.29 mmol), Xphos (8.6 mg, 0.018 mmol),tris(dibenzylideneacetone)dipalladium (4.7 mg, 0.0052 mmol) and3-bromo-2-methylpyridine (50 mg, 0.29 mmol) in a dry, sealed microwavetube under an atmosphere of argon was added tert-butyl alcohol (1.0 mL)followed by a 1.0 M solution of potassium tert-butoxide in tert-butylalcohol (0.58 mL, 0.58 mmol). The mixture was heated under microwaveirradiation at 150° C. for 30 min. The reaction mixture was allowed tocool to rt and then added to water (50 mL). The precipitated solid wascollected via suction filtration, washed with water, and dried toproduce9-chloro-10-[3-(dimethylamino)propyl]-2-[(2-methylpyridin-3-yl)amino]-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(95 mg, 0.21 mmol, 75%) as a brown solid. LCMS (FA): m/z=453.5 (M+H).

Compounds in the following table were prepared from the appropriatestarting materials using the procedures described above:

9-chloro-10-[3-(dimethylamino)propyl]-2-(pyridin-3-ylamino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one9-chloro-2-({5-[3-(dimethylamino)propyl]-2,6-dimethylpyridin-3-yl}amino)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

2-{[5-(3-Aminopropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

Step 1: tert-Butyl[3-(6-methyl-5-{[6-oxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridin-3-yl)propyl]carbamate

To a solution of(4Z)-4-[(dimethylamino)methylene]-8-(trifluoromethyl)-3,4-dihydro-1H-1-benzazepine-2,5-dione(1.05 g, 3.51 mmol) and tert-butyl[3-(5-{[amino(imino)methyl]amino}-6-methylpyridin-3-yl)propyl]carbamate(1.62 g, 5.27 mmol) in EtOH (50 mL) was added potassium carbonate (2.91g, 21.1 mmol). The reaction mixture was allowed to stir at 80° C.overnight. The reaction mixture was diluted with water (100 mL) andEtOAc (50 mL). The organic solution was separated and the aqueoussolution was extracted with EtOAc (2×50 mL). The organic solutions werecombined, washed with brine (100 mL), dried over MgSO₄, filtered andconcentrated to give tert-butyl[3-(6-methyl-5-{[6-oxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridin-3-yl)propyl]carbamate(2.13 g, >99%) as a dark brown solid. LCMS (FA): R_(t)=1.79 min,m/z=543.0 (M+H).

Step 2:2-{[5-(3-Aminopropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To a 4M solution of HCl in dioxane (20 mL) was added tert-butyl[3-(6-methyl-5-{[6-oxo-9-(trifluoromethyl)-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl]amino}pyridin-3-yl)propyl]carbamate(1.10 g, 2.04 mmol). The reaction mixture was allowed to stir at rtovernight and then concentrated to give2-{[5-(3-aminopropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(1.35 g, 2.30 mmol, >99%) as a brown solid. LCMS (FA): R_(t)=0.55 min,m/z=443.0 (M+H).

2-[(6-Aminopyridin-3-yl)amino]-9-chloro-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To a solution ofN-{5-[(9-chloro-6-oxo-6,7-dihydro-5H-pyrimido[5,4-d][1]benzazepin-2-yl)amino]pyridin-2-yl}-2,2-dimethylpropanamide(0.220 g, 0.38 mmol) in THF (3 mL) and MeOH (4 mL) at rt was added 6MHCl in water (5 mL). The solution was allowed to stir at 100° C. for 22h. The organic solvents were allowed to boil off and the aqueous mixturewas allowed to cool to rt. Solid K₂CO₃ was added and the mixture wasfiltered. The resulting solid was collected and purified by columnchromatography to give2-[(6-aminopyridin-3-yl)amino]-9-chloro-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.095 g, 71%).

2-{[5-[3-(Dimethylamino)propyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

Step 1:2-{[5-Chloro-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A mixture of2-amino-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.42 g, 1.44 mmol), 3-bromo-5-chloro-2-(trifluoromethyl)pyridine (0.37g, 1.44 mmol), Xphos (0.054 g, 0.115 mmol), andtris(dibenzylideneacetone)dipalladium(0) (0.052 g, 0.057 mmol) in asealed microwave tube was evacuated and purged with argon 3 times. Tothe vial were added tert-butyl alcohol (5.3 mL) and 1.00 M of t-BuOK intert-butyl alcohol (2.0 mL, 2.0 mol) via syringe. The mixture wasallowed to stir to mix well, then was subjected to microwave irradiationat 150° C. for 1200 sec. The mixture was allowed to cool to rt and THF(6 mL) was added. The mixture was filtered and the solid was washed withTHF. The organic solutions were combined and concentrated. The residuewas purified by column chromatography to give2-{[5-chloro-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.081 g, 12%) as a solid.

Step 2:2-[5-[3-(Dimethylamino)prop-1-yn-1-yl]-2-(trifluoromethyl)pyridin-3-yl]amino-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

A mixture of2-{[5-chloro-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.058 g, 0.12 mmol), propargyl(dimethylamine) (0.016 mL, 0.15 mmol) andcesium carbonate (0.080 g, 0.25 mmol) in DMF (0.60 ml) was degassed for15 min with argon. To the suspension were addedbis(acetonitrile)palladium(II) chloride (0.0020 g, 0.0061 mmol) andXphos (0.0076 g, 0.016 mmol). The suspension was purged three times withnitrogen and was then allowed to stir at 150° C. for 48 h. The mixturewas filtered through celite and the filtrate was concentrated. Theresidue was purified by column chromatography to give2-{[5-[3-(dimethylamino)prop-1-yn-1-yl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[1]benzazepin-6-one(0.042 g, 66%) as a solid.

Step 3:2-{[5-[3-(Dimethylamino)propyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To a round bottom flask were added2-{[5-[3-(dimethylamino)prop-1-yn-1-yl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.042 g, 0.081 mmol), EtOH (5 mL), THF (1 mL), and Raney nickel (20mg). The resulting mixture was allowed to stir under a hydrogen balloonovernight. The mixture was filtered through celite and the filtrate wasconcentrated. The residue was purified by column chromatography to give2-{[5-[3-(dimethylamino)propyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.033 g, 78%) as a solid.

Example 5 Separation of enantiomers of10-{[6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-3-(trifluoromethyl)-5,7-dihydro-6H-dibenzo[b,d]azepin-6-one

The enantiomers of10-{[6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]amino}-3-(trifluoromethyl)-5,7-dihydro-6H-dibenzo[b,d]azepin-6-onewere separated on a Vision HPLC fitted with a Chiralpak IC (20×250 mm)column using a isocratic solvent system of 80% hexane:10% iso-propylalcohol:10% ethanol:0.1% diethylamine at a flow rate of 20 mL/min.

Example 6 Synthesis of Compounds of Formula I

2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(I-111)

To a solution of2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(545 mg, 1.16 mmol) in pyridine (10 mL) was added phosphoruspentasulfide (810 mg, 3.6 mmol). The resulting suspension was allowed tostir at 60-65° C. for 18 h. The suspension was cooled to 10-15° C. and 2mL of water were added. The mixture was then allowed to stir at rt for5-10 min, during which time the solids dissolved. The solution was addedslowly to 1M aq. NaHCO₃ (80 ml) with stirring. The resulting mixture wasallowed to stir for 2 h and was then extracted with EtOAc (3×60 mL). Theorganic solutions were combined, washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified by HPLC togive2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneas the formate salt (378 mg, 61%) LCMS (FAP1): R_(t)=8.3 min., m/z=487.3(M+H).

The formate salt of2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(1.63 g, 3.06 mmol) was dissolved in water (50 ml) and the resultingsolution was added to a mixture of sat. NaHCO₃ (30 ml), 1M Na₂CO₃ (30ml) and EtOAc with vigorous stirring. Brine (30 ml) was added and themixture was allowed to stir for 15 min. The aqueous solution wasseparated and extracted with EtOAc (2×30 ml). The organic solutions werecombined, washed with brine (2×40 ml), dried over Na₂SO₄, filtered andconcentrated to give2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(1.41 g, 95%).

A solution of2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thionein a mixture of MeOH (1.6 ml) and MeCN (10 ml) was allowed to stir undergentle reflux. To this solution was added 12 M HCl (15 ul, 0.19 mmol).The solution was seeded and allowed to cool slowly to rt. The resultingslurry was left at rt for 18 hrs. The solid was collected by filtrationand dried under high vacuum to give the monohydrochloride salt of2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(69 mg, 71%).

2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]-benzazepine-6-thione(I-111)

A mixture of2-({5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(175 g, 372 mmol), phosphorus pentasulfide (82.7 g, 372 mmol) andpyridine (1.35 L) was heated at 50° C. for 18 hours. HPLC analysisshowed completion of the reaction. After the reaction mixture was cooledbelow room temperature, a solution of sodium carbonate (525 g, 4.95 mol)in 5.1 L of water was slowly added while the temperature was kept atbelow room temperature. The resulting biphasic solution was separatedand the organic layer was washed with the same amount of sodiumcarbonate solution two times. The organic phase was then reduced to ca.1.1 L in volume. The solution was diluted with water (1.0 L) and stirredat room temperature overnight resulting in a yellow suspension. Thesolid was filtered and washed with 200 mL EtOH/water (1 vol/2vol), andwater 200 mL. After drying overnight under vacuo at 40° C., the crudeproduct was dissolved in 1.2 L of EtOH at reflux. Water (2.0 L) wasslowly added at 75° C. to induce crystallization. The suspension wasthen slowly cooled to room temperature and stirred overnight. The solid2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thionewas filtered and dried under vac at 40° C. to a constant weight (140.0g, HPLC purity >98%, 77% yield). LCMS (FA): R_(t)=6.26 min, m/z=485.2(M−H).

Compounds in the following table may be prepared from the appropriatestarting materials as either the formate or hydrochloride salt using theprocedures described above:

I-53 LCMS (FAL): R_(t) = 4.60 min, m/z = 493.6 (M + H). I-63 LCMS (FAL):R_(t) = 5.07 min, m/z = 527.7 (M + H). I-44 LCMS (FAP1): R_(t) = 7.56min, m/z = 479.6 (M + H). I-24 LCMS (FAP1): R_(t) = 7.66 min, m/z =469.5 (M + H). I-27 LCMS (FAP1): R_(t) = 7.23 min, m/z = 457.5 (M + H).I-8  LCMS (FAP1): R_(t) = 7.43 min, m/z = 469.4 (M + H). I-93 LCMS (FA):R_(t) = 6.60 min, m/z = 453.5 (M + H). I-33 LCMS (FA): R_(t) = 4.95 min,m/z = 439.4 (M + H). I-61 LCMS (FA): R_(t) = 4.95 min, m/z = 486.2 (M +H). I-13 LCMS (FA): R_(t) = 5.59 min, m/z = 439.2 (M + H). I-28 LCMS(FA): R_(t) = 4.55 min, m/z = 452.2 (M + H). I-71 LCMS (FA): R_(t) =8.10 min, m/z = 468.6 (M + H).  I-102 LCMS (FA): R_(t) = 12.04 min, m/z= 453.4 (M + H). I-47 LCMS (FA): R_(t) = 7.30 min, m/z = 423.4 (M + H).I-83 LCMS (FA): R_(t) = 4.05 min, m/z = 538.7 (M + H). I-80 LCMS (FAP1):R_(t) = 8.0 min, m/z = 486 (M + H). I-5  LCMS (AAL): R_(t) = 7.5 min,m/z = 447 (M + H). I-79 LCMS (FAP1): R_(t) = 4.5 min, m/z = 444 (M + H).I-60 LCMS (FAP1): R_(t) = 3.6 min, m/z = 432 (M + H). I-39 LCMS (FAP1):R_(t) = 4.0 min, m/z = 459 (M + H). I-32 LCMS (FA): R_(t) = 6.02 min,m/z = 473.4 (M + H). I-90 LCMS (FAL): R_(t) = 4.02 min, m/z = 459.1 (M +H). I-22 LCMS (FA): R_(t) = 3.54 min, m/z = 480.4 (M + H). I-1  LCMS(FAP1): R_(t) = 3.7 min, m/z = 449 (M + H). I-91 LCMS (FAL): R_(t) =3.91 min, m/z = 439.2 (M + H).  I-108 LCMS (FAL): R_(t) = 3.38 min, m/z= 453.2 (M + H). I-50 LCMS (FAL): R_(t) = 4.01 min, m/z = 473 (M + H).I-99 LCMS (FAP1): R_(t) = 7.46 min, m/z = 437.5 (M − H). I-26 LCMS(FAL): R_(t) = 5.28 min, m/z = 528.7 (M + H). I-6  LCMS (FAL): R_(t) =5.28 min, m/z = 528.7 (M + H). I-87 LCMS (FAP1): R_(t) = 8.91 min, m/z =513.7 (M + H). I-54 LCMS (FAP1): R_(t) = 7.99 min, m/z = 479 (M + H).I-72 LCMS (FAP1): R_(t) = 7.61 min, m/z = 482 (M + H). I-43 LCMS (FAP1):R_(t) = 8.53 min, m/z = 516 (M + H). I-30 LCMS (FAL): R_(t) = 5.97 min,m/z = 479.5 (M + H). I-34 LCMS (FAL): R_(t) = 3.48 min, m/z = 522.1 (M +H). I-25 LCMS (FAP1): R_(t) = 5.96 min, m/z = 460 (M + H).  I-104 LCMS(FAP1): R_(t) = 5.49 min, m/z = 425 (M + H). I-78 LCMS (AAL): R_(t) =6.83 min, m/z = 467.3 (M + H). I-51 LCMS (AAL): R_(t) = 5.99 min, m/z =480.4 (M + H). I-45 LCMS (FAP1): R_(t) = 7.66 min, m/z = 465 (M + H).I-89 LCMS (FAL): R_(t) = 3.62 min, m/z = 467 (M + H). I-12 LCMS (FAL):R_(t) = 3.84 min, m/z = 501 (M + H). I-52 LCMS (FAL): R_(t) = 3.60 min,m/z = 556.3 (M + H). I-59 LCMS (FAP1): R_(t) = 5.56 min, m/z = 522.5(M + H). I-15 LCMS (FAP1): R_(t) = 5.92 min, m/z = 508.5 (M + H). I-29LCMS (FAP1): R_(t) = 8.90 min, m/z = 528.6 (M + H). I-94 LCMS (FAP1):R_(t) = 5.15 min, m/z = 453 (M + H). I-95 LCMS (FAP1): R_(t) = 10.84min, m/z = 514.6 (M + H). I-3  LCMS (FAP1): R_(t) = 9.70 min, m/z =480.5 (M + H). I-69 LCMS (FAP1): R_(t) = 7.88 min, m/z = 494.5 (M + H).I-85 LCMS (FAP1): R_(t) = 6.95 min, m/z = 466.5 (M + H). I-23 LCMS(FAP1): R_(t) = 5.47 min, m/z = 502 (M + H). I-56 LCMS (FAP1): R_(t) =7.23 min, m/z = 481.3 (M + H). I-76 LCMS (FAP1): R_(t) = 3.84 min, m/z =469 (M + H). I-88 LCMS (FAP1): R_(t) = 8.60 min, m/z = 516.6 (M + H).I-41 LCMS (FAP1): R_(t) = 7.83 min, m/z = 482.5 (M + H). I-92 LCMS (FA):R_(t) = 3.88 min, m/z = 515 (M + H). I-7  LCMS (FAP1): R_(t) = 9.72 min,m/z = 494.5 (M + H). I-19 LCMS (FAP2): R_(t) = 6.94 min, m/z = 467.6(M + H). I-9  LCMS (FAP1): R_(t) = 8.90 min, m/z = 528.5 (M + H). I-75LCMS (FAP1): R_(t) = 3.93 min, m/z = 528 (M + H). I-57 LCMS (FAP1):R_(t) = 7.77 min, m/z = 459.6 (M + H). I-18 LCMS (FAP1): R_(t) = 7.44min, m/z = 501.6 (M + H).  I-105 LCMS (FAP1): R_(t) = 7.66 min, m/z =494.6 (M + H). I-64 LCMS (FAP1): R_(t) = 8.51 min, m/z = 488.6 (M + H).I-97 LCMS (FAP2): R_(t) = 5.21 min, m/z = 454.6 (M + H).  I-112 LCMS(FA): R_(t) = 4.62 min, m/z = 468 (M + H). I-35 LCMS (FAP1): R_(t) =7.25 min, m/z = 502.6 (M + H). I-81 LCMS (FAP1): R_(t) = 10.26 min, m/z= 382.5 (M + H). I-4  LCMS (FAP1): R_(t) = 7.27 min, m/z = 416.4 (M +H).  I-100 LCMS (FAP1): R_(t) = 8.95 min, m/z = 487.4 (M + H). I-77 LCMS(FAP1): R_(t) = 7.74 min, m/z = 515.6 (M + H).  I-101 LCMS (FAP1): R_(t)= 8.53 min, m/z = 487.5 (M + H). I-98 LCMS (FAP1): R_(t) = 7.14 min, m/z= 416.4 (M + H). I-16 LCMS (FAP1): R_(t) = 7.62 min, m/z = 453.4 (M +H). I-37 LCMS (FAP3): R_(t) = 4.38 min, m/z = 382.2 (M + H). I-82 LCMS(FAP1): R_(t) = 8.10 min, m/z = 451.5 (M + H).  I-110 LCMS (FAP1): R_(t)= 7.69 min, m/z = 471.4 (M + H).  I-106 LCMS (FAP1): R_(t) = 11.24 min,m/z = 437.6 (M + H). I-73 LCMS (FAP1): R_(t) = 9.91 min, m/z = 466.5(M + H).  I-109 LCMS (FAP1): R_(t) = 8.09 min, m/z = 453.5 (M + H). I-14LCMS (FAP1): R_(t) = 7.08 min, m/z = 465.6 (M + H). I-84 LCMS (FAP1):R_(t) = 7.42 min, m/z = 497.2 (M + H). I-17 LCMS (FAP1): R_(t) = 3.62min, m/z = 479.1 (M + H). I-38 LCMS (FAP1): R_(t) = 5.47 min, m/z =467.4 (M + H). I-42 LCMS (FAP1): R_(t) = 6.89 min, m/z = 467.5 (M + H).I-62 LCMS (FAL): R_(t) = 4.06 min, m/z = 473.0 (M + H). I-67 LCMS (FAL):R_(t) = 4.09 min, m/z = 500.4 (M + H). I-40 LCMS (FAL): R_(t) = 3.57min, m/z = 494 (M + H). I-65 LCMS (FAL): R_(t) = 3.90 min, m/z = 528.4(M + H). I-74 LCMS (FAL): R_(t) = 3.96 min, m/z = 512.4 (M + H). I-46LCMS (FA): R_(t) = 1.38 min, m/z = 500.2 (M + H).  I-103 LCMS (FAP1):R_(t) = 3.64 min, m/z = 466.1 (M + H). I-66 LCMS (FAP1): R_(t) = 3.77min, m/z = 494.0 (M + H). I-49 LCMS (FAP1): R_(t) = 3.82 min, m/z =508.0 (M + H). I-2  LCMS (FAP1): R_(t) = 3.91 min, m/z = 528.0 (M + H).I-58 LCMS (FAP1): R_(t) = 7.22 min, m/z = 480.5 (M + H).  I-107 LCMS(FAP1): R_(t) = 7.56 min, m/z = 452.0 (M + H). I-11 LCMS (FAP1): R_(t) =8.06 min, m/z = 480.4 (M + H).

9-Chloro-2-{[5-[2-(dimethylamino)ethyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(I-70)

To a solution of9-chloro-2-{[5-[2-(dimethylamino)ethyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.011 g, 0.023 mmol) in THF (1.5 ml) was added Lawesson's reagent(0.028 g, 0.069 mmol). The resulting mixture was allowed to stir for 3 hat 65° C. and then allowed to cool to rt. HCl (6 mL, 1M in ether) wasadded. The mixture was filtered and the solid was washed with THF andsaturated NaHCO₃. The residue was purified by column chromatography togive9-chloro-2-{[5-[2-(dimethylamino)ethyl]-2-(trifluoromethyl)pyridin-3-yl]amino}-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(I-70) as the formate salt (7.0 mg, 56%). LCMS (FA): R_(t)=7.10 min,m/z=493.1 (M+H).

Compounds in the following table were prepared from the appropriatestarting materials as either the formate or hydrochloride salt using theprocedures described above:

I-68 LCMS (FA): R_(t) = 4.40 min, m/z = 369.2 (M + H). I-96 LCMS (FA):R_(t) = 6.92 min, m/z = 465.6 (M + H). I-36 LCMS (FA): R_(t) = 5.74 min,m/z = 541.6 (M + H). I-10 LCMS (FAP1): R_(t) = 8.19 min, m/z = 509.5(M + H). I-31 LCMS (FAL): R_(t) = 4.22 min, m/z = 509.2 (M + H). I-86LCMS (FA): R_(t) = 1.23 min, m/z = 471.3 (M + H). I-114 LCMS (FAL):R_(t) = 7.79 min, m/z = 499.5 (M + H). I-113 LCMS (FAL): R_(t) = 7.79min, m/z = 499.1 (M + H). I-21 LCMS (FAP1): R_(t) = 7.56 min, m/z =558.6 (M + H).

Example 6 Synthesis of2-{[5-(3-Hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(I-55)

Step 1:2-{[5-(3-{[tert-Butyl(dimethyl)silyl]oxy}propyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

To a solution of2-{[5-(3-hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.25 g, 0.564 mmol) in THF (10 ml) were added imidazole (0.077 g, 1.13mmol) and tert-butyldimethylsilyl chloride (0.10 g, 0.676 mmol). Thereaction mixture was allowed to stir overnight at rt and was thenconcentrated. Water (10 ml) and saturated NaHCO₃ (10 ml) were added. Theresulting mixture was allowed to stir for 1 h at rt. The mixture wasextracted with EtOAc (2×30 mL). The organic solutions were combined,dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby column chromatography to give2-{[5-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.16 g, 0.29 mmol, 51%).

Step 2:2-{[5-(3-Hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(I-55)

To a solution of2-{[5-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one(0.16 g, 0.29 mmol) in THF (7 mL) was added2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,3,2,4-dithiadiphosphetane (0.23g, 0.574 mmol). The resulting mixture was allowed to stir for 6 h at rt.The mixture was concentrated. Water (20 mL) and saturated NaHCO₃ (20 mL)were added. The mixture was extracted with DCM (2×30 mL). The organicsolutions were combined, dried over Na₂SO₄, filtered and concentrated.The residue was purified by column chromatography to give2-{[5-(3-hydroxypropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneas the formate salt (0.055 g, 0.11 mmol, 38%) as a solid. LCMS (FA):R_(t)=5.10 min, m/z=460.3 (M+H).

Example 7 Synthesis of2-{[5-(3-aminopropyl)-2-methylpyridin-3-yl]amino}-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepin-6-one

Step 1: Diethyl (5-bromo-3-nitropyridin-2-yl)malonate

To a suspension of NaH (60% in mineral oil, 27.9 g, 0.69 mol) in DMF(300 mL) at 5-10° C. was slowly added ethyl malonate (125 mL, 0.69 mol)over 30 min. The mixture was allowed to stir for 20 min at rt, duringwhich time the suspension became a solution. A solution of5-bromo-2-chloro-3-nitropyridine (75 g, 0.32 mol) in DMF (75 mL) wasadded slowly at 5-10° C. The resulting dark red mixture was allowed tostir at 40° C. for 2 h. The reaction mixture was then poured into 1MAcOH (0.75 L) and extracted with DCM (3×250 mL). The organic solutionswere combined, washed with water and brine, dried over MgSO₄, filteredand concentrated. The residue was purified by column chromatography togive diethyl (5-bromo-3-nitropyridin-2-yl)malonate (260 g, 99%) as ayellow oil.

Step 2: 5-Bromo-2-methyl-3-nitropyridine

To diethyl (5-bromo-3-nitropyridin-2-yl)malonate (66 g, 0.18 mol) wasadded water (250 mL) and 12 M HCl (360 mL, 4.32 mol). The mixture washeated at 105° C. until TLC showed starting material was consumed. Thereaction mixture was then allowed to cool to rt and brine (0.67 L) wasadded. The organic solution was separated and the aqueous solution wasextracted with DCM (3×0.67 L). The organic solutions were combined,washed with brine, sat. aq. NaHCO₃, and brine again, dried over Na₂SO₄,filtered and concentrated to give 5-bromo-2-methyl-3-nitropyridine (34.7g, 75%) as a yellow solid.

Compounds in the following table were prepared from the appropriatestarting materials using the procedures described above:

5-bromo-2-ethyl-3-nitropyridine

Example 8 Synthesis of Guanidines of Formula III Used for thePreparation of Compounds of Formula vi (Formula IV)

N-[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]guanidineStep 1: tert-Butyl-3,3-dimethyl-4-oxopiperidine-1-carboxylate

To a solution of tert-butyl 4-oxopiperidine-1-carboxylate (15.0 g, 75.3mmol) in THF (400 mL) at 0° C. was added sodium hydride (3.63 g, 151mmol) slowly. The reaction mixture was allowed to stir for 5 min andthen methyl iodide (11.7 mL, 188 mmol) was added dropwise. The reactionmixture was allowed to stir for 30 min at 0° C. and at rt overnight. Thereaction mixture was diluted with water (200 mL). The aqueous solutionwas extracted with DCM. The organic solutions were combined, washed withbrine, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography to givetert-butyl-3,3-dimethyl-4-oxopiperidine-1-carboxylate (7.0 g, 40%) as awhite solid. LCMS (FA): R_(t)=1.77 min, m/z=228.3 (M+H).

Step 2:tert-Butyl-3,3-dimethyl-4-{[(trifluoromethyl)sulfonyl]oxy}-3,6-dihydropyridine-1(2H)-carboxylate

To a solution of tert-butyl 3,3-dimethyl-4-oxopiperidine-1-carboxylate(2.18 g, 9.59 mmol) in THF (20 mL) was added lithiumbis(trimethylsilyl)amide (1.0M in THF, 10.5 mL) dropwise at −78° C. Thereaction mixture was allowed to stir for 30 min and thenN-phenylbis(trifluoromethanesulphonimide (3.70 g, 10.4 mmol) in THF (10mL) was added dropwise. The reaction mixture was allowed to warm to 0°C. then rt, and allowed to stir overnight. The mixture was quenched bythe addition of sat. aq. NH₄Cl (40 mL) and brine (80 mL). After stirringfor 30 min, the aqueous solution was extracted with DCM. The organicsolutions were combined, dried over Na₂SO₄, filtered and concentrated.The residue was purified by column chromatography to givetert-butyl-3,3-dimethyl-4-{[(trifluoromethyl)sulfonyl]oxy}-3,6-dihydropyridine-1(2H)-carboxylate(2.3 g, 67%). LCMS (FA): R_(t)=2.34 min, m/z=360.1 (M+H).

Step 3:tert-Butyl-3,3-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate

To a solution of tert-butyl3,3-dimethyl-4-{[(trifluoromethyl)sulfonyl]-oxy}-3,6-dihydropyridine-1(2H)-carboxylate(1.55 g, 4.31 mmol) in 1,4-dioxane (30 mL) were addedbis(pinacolato)diboron (1.20 g, 4.74 mmol),1,1′-bis(diphenylphosphino)ferrocenepalladium (II) dichloride,dichloromethane (106 mg, 0.129 mmol),1,1′-bis(diphenylphosphino)ferrocene (71.7 mg, 0.129 mmol) and potassiumacetate (1.27 g, 12.9 mmol). The reaction mixture was flushed withnitrogen gas and allowed to stir for 5 h at 80° C. The reaction mixturewas filtered through Celite® and the filter cake was washed with EtOAc.The filtrate was washed with brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to givetert-butyl-3,3-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(0.55 g, 38%) as a white solid. LCMS (FA): R_(t)=2.51 min, m/z=338.3(M+H).

Step 4:tert-Butyl-3′,3′,6-trimethyl-5-nitro-3′,6′-dihydro-3,4′-bipyridine-1′(2′H)-carboxylate

To a solution oftert-butyl-3,3-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate(2.12 g, 6.28 mmol) in 1,4-dioxane (90 mL) were added5-bromo-2-methyl-3-nitropyridine (1.64 g, 7.54 mmol),tetrakis(triphenylphosphine) palladium (642 mg, 0.556 mmol), potassiumphosphate (4.96 g, 23.4 mmol) and water (22 mL). The reaction mixturewas flushed with nitrogen gas and allowed to stir for 24 h at 80° C. Thereaction mixture was filtered through a Celite® pad and washed with DCM.The filtrate was washed with brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to givetert-butyl-3′,3′,6-trimethyl-5-nitro-3′,6′-dihydro-3,4′-bipyridine-1′(2′H)-carboxylate(1.3 g, 60%) as a white solid. LCMS (FA): R_(t)=2.16 min, m/z=348.3(M+H).

Step 5: 3′,3′,6-Trimethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine[HCl]

To a solution oftert-butyl-3′,3′,6-trimethyl-5-nitro-3′,6′-dihydro-3,4′-bipyridine-1′(2′H)-carboxylate(1.30 g, 3.70 mmol) in MeOH (30 mL) was added hydrochloric acid (4.0 Min 1,4-dioxane, 40 mL). The reaction mixture was allowed to stir for 2 hat rt. The reaction mixture was concentrated to give3′,3′,6-trimethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine as thehydrochloride salt (1.12 g, 100%) as a white solid. LCMS (FA):R_(t)=0.89 min, m/z=248.3 (M+H).

Step 6:1′,3′,3′,6-Tetramethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine

To a solution of3′,3′,6-trimethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine [HCl](1.12 g, 3.50 mmol) in DCM (55 mL) were added formaldehyde (37% inwater, 4.0 mL) and sodium triacetoxyborohydride (4.45 g, 21.0 mmol). Thereaction mixture was allowed to stir for 2 h at rt. The mixture wasquenched by the addition of sat. aq. sodium bicarbonate (40 mL). Afterstirring for 10 min, the aqueous solution was extracted with DCM. Theorganic solutions were combined, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to give1′,3′,3′,6-tetramethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine(0.914 g, 100%). LCMS (FA): R_(t)=0.85 min, m/z=262.4 (M+H).

Step 7: 2-Methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-amine

To a solution of1′,3′,3′,6-tetramethyl-5-nitro-1′,2′,3′,6′-tetrahydro-3,4′-bipyridine(0.912 g, 3.49 mmol) in MeOH (70 mL) was added palladium hydroxide (20wt. % Pd (dry basis) on carbon, 808 mg). The reaction mixture wasflushed with hydrogen gas and allowed to stir at rt overnight under anatmosphere of hydrogen. The reaction mixture was filtered through aCelite® pad and washed with MeOH. The filtrate was concentrated to give2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-amine (0.814 g,100%). LCMS (FA): R_(t)=0.24 min, m/z=234.4 (M+H).

Step 8: Di-tert-butyl((Z)-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]amino}methylylidene)biscarbamate

To a solution of2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-amine (0.814 g,3.49 mmol) in DCM (50 mL) were added1,3-bis(tert-butoxycarbonyl)-2-methyl-2-thiopseudourea (1.52 g, 5.23mmol), mercury (II) chloride (1.42 g, 5.23 mmol) and TEA (2.92 mL, 20.9mmol). The reaction mixture was allowed to stir overnight at rt thenfiltered through a Celite® pad and washed with DCM. The filtrate waswashed with brine, dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by column chromatography to give di-tert-butyl((Z)-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]amino}methylylidene)biscarbamate(0.79 g, 48%) as a white solid. LCMS (FAL): R_(t)=5.08 min, m/z=476.6(M+H).

Step 9:N-[2-Methyl-5-(1,3,3-trimethyl)piperidin-4-yl)pyridine-3-yl]guanidine

To a solution of di-tert-butyl((Z)-{[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]amino}methylylidene)biscarbamate(0.78 g, 1.60 mmol) in DCM (10 mL) was added hydrochloric acid (4.0 M in1,4-dioxane, 20 mL). The reaction mixture was allowed to stir at rtovernight. The reaction mixture was concentrated to give thehydrochloride salt ofN-[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]guanidine(750 mg, 110%) as a white solid.

Steps 7-9 in the synthesis ofN-[2-methyl-5-(1,3,3-trimethylpiperidin-4-yl)pyridine-3-yl]guanidinerepresent General Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-[2-methyl-5-(1-methylpiperidin-4-yl)pyridin-3-yl]guanidineN-[5-(1-ethylpiperidin-4-yl)-2-methylpyridin-3-yl]guanidineN-{5-[1-(2-fluoroethyl)piperidin-4-yl]-2-methylpyridin-3-yl}guanidine

N-{2-methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridine-3-yl}guanidine

Step 1: 2-Methyl-6-[(1-methylpiperidin-4-yl)-methyl]-3-nitropyridine

To a solution of 9-BBN in THF (0.5 M, 4.93 mL, 2.46 mmol) was added1-methyl-4-methylenepiperidine (274 mg, 2.46 mmol). The reaction mixturewas allowed to stir at 75° C. for 1 h and was then allowed to cool tort. The resulting solution was transferred into a solution of2-methyl-5-nitro-6-methylpyridine (486 mg, 2.24 mmol) in DMF (11 mL). Tothe mixture were added[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (1:1complex with DCM, 54.9 mg, 0.0672 mmol) and potassium carbonate (402 mg,2.91 mmol). The reaction mixture was allowed to stir for 3 h at rt. Themixture was quenched by the addition of water (10 mL). The aqueoussolution was extracted with DCM. The organic solutions were combined,washed with brine, dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by column chromatography to give2-methyl-6-[(1-methylpiperidin-4-yl)-methyl]-3-nitropyridine (64 mg,10%) as a white solid. LCMS (FAL): R_(t)=3.38 min, m/z=250.2 (M+H).

Step 2:N-{2-methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridine-3-yl}guanidine

The hydrochloride salt ofN-{2-methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridine-3-yl}guanidinewas prepared as a white solid from2-methyl-6-[(1-methylpiperidin-4-yl)-methyl]-3-nitropyridine followingthe procedures of General Method 1.

N-{5-[3-(Dimethylamino)propyl]-2-methoxypyridin-3-yl}guanidine

Step 1: 3-(6-Methoxy-5-nitropyridin-3-yl)-N,N-dimethylprop-2-yn-1-amine

To a solution of 5-bromo-2-methoxy-3-nitropyridine (1.50 g, 6.44 mmol)in DMF (30 mL) were added bis(acetonitrile)palladium (II) chloride (83.5mg, 0.322 mmol),2-dicyclohexylphosphino-2′,4′,6′-tri-1-propyl-1-1′-biphenyl (399 mg,0.837 mmol), cesium carbonate (4.19 g, 12.9 mmol) andpropargyl(dimethylamine) (0.832 mL, 7.72 mmol). The reaction mixture wasflushed with nitrogen gas and allowed to stir for 4 h at 50° C. Thereaction mixture was diluted with EtOAc and washed with brine, driedover Na₂SO₄, filtered and concentrated. The residue was purified bycolumn chromatography to give3-(6-methoxy-5-nitropyridin-3-yl)-N,N-dimethylprop-2-yn-1-amine (0.41 g,27%) as a white solid. LCMS (FA): R_(t)=0.85 min, m/z=236.2 (M+H).

Step 2:N-{2-Methyl-6-[(1-methylpiperidin-4-yl)methyl]pyridine-3-yl}guanidine

N-{2-Methyl-6[(1-methylpiperidin-4-yl)methyl]pyridine-3-yl}guanidine wasprepared as the hydrochloride salt as a white solid from3-(6-methoxy-5-nitropyridin-3-yl)-N,N-dimethylprop-2-yn-1-aminefollowing the procedures of General Method 1.

N-{5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}guanidine

Step 1: N,N-Dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine

To a mixture of 5-bromo-2-methyl-3-nitropyridine (6.50 g, 30.0 mmol),copper iodide (280 mg, 1.5 mmol) andbis(triphenylphosphine)palladium(II) chloride (520 mg, 0.75 mmol) indiethylamine (17 mL) was added propargyl(dimethylamine) (4.04 mL, 37.5mmol). The reaction mixture was allowed to stir for 15 h at rt. Theresulting mixture was diluted with EtOAc (200 mL) and 1M aqueous Na₂CO₃(150 mL). After stirring for 30 min, the organic solution was separatedand the aqueous solution was extracted with EtOAc (2×100 mL). Theorganic solutions were combined, washed with water and brine, dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography to giveN,N-dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine (5.98 g,91%) as a dark oil. LCMS (FA): R_(t)=0.77 min, m/z=220.1 (M+H).

Step 2: 5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-amine

To a solution ofN,N-dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine (3.18 g,13.8 mmol) in EtOH (50 mL) under an atmosphere of N₂ was added Raney2800 nickel (50% water slurry; 760 mg, 4 mmol) using 5 mL of ethanol totransfer. The mixture was purged with H₂ (1 atm) and allowed to stir atrt under an atmosphere of H₂ for 20 h. More Raney nickel (620 mg, 3.6mmol) was added using 5 mL of ethanol to transfer. The mixture waspurged with H₂ (1 atm) and allowed to stir at rt under an atmosphere ofH₂ for 20 h. The mixture was filtered through Celite® and the filtratewas concentrated to give5-[3-(dimethylamino)propyl]-2-methylpyridin-3-amine (2.45 g, 93%) as abrown solid. LCMS (AAL): R_(t)=0.98 min, m/z=194.4 (M+H).

Step 3: N-{5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}guanidine

The hydrochloride salt ofN-{5-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}guanidine wasprepared from 5-[3-(dimethylamino)propyl]-2-methylpyridin-3-aminefollowing General Method 1, with the exception that the hydrogenationstep was performed using Raney Ni as described in Step 2. LCMS (AAL):R_(t)=0.82 min, m/z=236.6 (M+H).

N-{5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}guanidinetrihydrochloride

Step 1: N,N-dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine

A 12 L reactor equipped with an overhead stirrer and N₂ bubbler wascharged with a solution of 5-bromo-2-methyl-3-nitropyridine (945 g, 4.35mol) in EtOAc (5.7 L, 6 vol). To the reaction mixture, Pd(PPh₃)₂Cl₂(15.4 g, 0.022 mol, 0.005 equiv), CuI (8.28 g, 0.0435 mol, 0.01 equiv)and Et₃N (3 L, 21.75 mol, 5 equiv) were added. Nitrogen was bubbledthrough the solution for 15 min and 3-dimethylamino-1-propyne (556 mL,5.22 mol, 1.2 equiv) was added. The reaction-temperature setting wasincreased to 60° C. stepwise. The maximum temperature attained was 66°C. The reaction was continued for 3 h at this temperature (60° C.) andwas judged complete by HPLC analysis. The reaction was cooled to ambienttemperature and the inorganic salts were filtered off by passing thereaction mixture through a pad of celite. The filter cake was washedwith EtOAc (2 L, 2 vol). The filtrate was transferred into a 20 Lseparatory funnel. The EtOAc layer was washed with 5% aqueous Na₂CO₃solution (2×5 L, 5 vol). The layers were separated. The EtOAc layer wasthen washed with 10% brine in water (5 L, 5 vol). The layers wereseparated and the EtOAc layer was pumped down to a low volume (ca. 2 L,2 vol) under reduced pressure. IPA (5 L, 5 vol) was added to thesolution and the solvent was again evaporated to a low volume (ca. 2 L,2 vol). The solution ofN,N-dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine wassubjected to hydrogenation in two batches using the procedure describedin step 2, below.

Step 2: 5-(3-(dimethylamino)propyl)-2-methylpyridin-3-aminehydrochloride

The solution ofN,N-dimethyl-3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-amine (2.17mol) in IPA (ca. 1 L) obtained in the previous step was diluted with IPAto make the total volume 2.3 L, and MeOH (2.3 L, 5 vol) was added to thesolution. This was then transferred into an 8 L Parr reactor. Thereactor was purged with N₂. The catalyst [Pd(OH)₂, 20 wt % on carbon wetwith 50% water, 15 wt %, 72 g] was charged into the reactor as a slurryin IPA (500 mL). The chiller temperature was set at 20° C. The reactorwas purged with N₂ (twice) and then with H₂ (twice). The pressure wasset at 30 psi. The internal temperature increased up to 62° C. even withthe chiller temperature set at 20° C. After the exotherm subsided andthe temperature cooled down to 35° C. (2 h), the chiller temperature wasincreased to 30° C. The reaction was continued at this temperature for 1h. No additional exotherm was observed. The chiller temperature wasincreased to 40° C. and the reaction was continued for 1 h. No exothermwas observed. Finally, the chiller temperature was set at 55° C.(internal temperature was 48° C.). Next, the pressure was ramped up to100 psi in increments of 10 psi over a period of 2 h. The reaction wascontinued for 20 h when ¹H NMR analysis indicated complete conversion.The reaction was cooled to ambient temperature and the catalyst wasfiltered off. The filtrate was used for the HCl salt formation.

The filtrates from Batch 1 and Batch 2 were combined. The combinedfiltrates were pumped down to a low volume (ca. 2 L). IPA (10 L, 10 vol)was added and the solvent was evaporated to a low volume (ca. 2 L). IPAc(2 L) was added and the resulting solution was cooled in an ice-waterbath. A solution of 5.95 N HCl in IPA (585 mL, 3.48 mol, 0.8 equiv) wasadded dropwise maintaining the internal temperature below 15° C. Thesuspension that resulted was stirred at ice-water bath temperature for 3h. The solid was collected by filtration and the filter cake was washedwith 1:1 IPA/IPAc (2 L). The solid was dried to a constant weight in avacuum oven at 35° C. to obtain5-(3-(dimethylamino)propyl)-2-methylpyridin-3-amine hydrochloride [620g, 62% yield over two steps] as an off-white solid. Solid separated outfrom the filtrate after letting it stand overnight at ambienttemperature. The solid was collected by filtration to obtain a secondcrop of 5-(3-(dimethylamino)propyl)-2-methylpyridin-3-aminehydrochloride [40 g, 4% recovery] as an off-white solid. The combinedyield was 660 g (66% yield over two steps). ¹H NMR (500 MHz, CD₃OD) δppm: 7.63 (s, 1H), 7.05 (s, 1H), 3.12-3.09 (m, 2H), 2.85 (s, 6H), 2.63(t, 2H, J=7.5 Hz), 2.34 (s, 3H), 2.05-1.98 (m, 2H); ESI-MS m/z: 194(M+H, 100%).

Step 3: N{5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}guanidinetrihydrochloride

A 1 L, three-neck, round-bottom flask equipped with an overhead stirrer,a reflux condenser, an N₂ inlet tube, and a temperature probe wascharged with 5-(3-(dimethylamino)propyl)-2-methylpyridin-3-aminehydrochloride (25 g, 0.109 mol). Acetonitrile (275 mL, 11 vol) was addedto it followed by acetic acid (25 mL, 1 vol). The initial suspensionturned into a clear solution after the addition of acetic acid.Chloroformamidine hydrochloride (15.6 g, 0.136 mol, 1.25 equiv) wasadded to the solution resulting in a suspension. The reaction was heatedat 40° C. for 30 min and at 50° C. for another 30 min. Then thetemperature was raised to 60° C. and the reaction was continued at thistemperature. After 1-2 h at 60° C., the reaction mixture became turbidand after 3-4 h, solid started separating out. The reaction wascontinued at 60° C. for 20 h and was judged complete by LC-MS analysis.The reaction was cooled to ambient temperature and the solid wascollected by filtration (N₂ flow was used during the filtration). Thefilter cake was washed with acetonitrile (50 mL, 2 vol). The solid wasdried under high vacuum overnight to obtainN-{5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}guanidinetrihydrochloride (32.1 g, 85% recovery) as a light brown solid.Elemental analysis Calcd: C, 41.81; H, 7.02; Cl, 30.85; N, 20.32. Found:C, 41.09; H, 6.72; Cl, 31.51; N, 20.47. ¹H NMR (500 MHz, DA)) δ ppm:8.57 (s, 1H), 8.45 (s, 1H), 3.22-3.19 (m, 2H), 2.90 (t, 2H, J=10 Hz),2.86 (s, 6H), 2.69 (s, 3H), 2.15-2.07 (m, 2H); ESI-MS m/z 236 (M+H,100%).

Compounds in the following table may be prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-{6-[3-(dimethylamino)propyl]-2-methylpyridin-3-yl}guanidineN-{5-[3-(dimethylamino)propyl]-6-methylpyridin-3-yl}guanidineN-{5-[3-(dimethylamino)propyl]-2-ethylpyridin-3-yl}guanidineN-{2-methyl-5-[3-(methylamino)propyl]pyridin-3-yl}guanidineN-[5-(3-hydroxypropyl)-2-methylpyridin-3-yl]guanidineN-{5-[3-(dimethylamino)propyl]pyridin-3-yl}guanidine

N-(2-Methyl-6-morpholin-4-ylpyridin-3-yl)guanidine

Step 1: 4-(6-Methyl-5-nitropyridin-2-yl)morpholine

To a solution of 2-fluoro-5-nitro-6-picoline (3.0 g, 19.2 mmol) in DMSO(26 mL) were added morpholine (3.35 mL, 38.4 mmol) and potassiumcarbonate (6.64 g, 48.0 mmol). The reaction mixture was allowed to stirovernight, at 70° C. The reaction mixture was then partitioned betweenEtOAc (40 mL) and water (40 mL). The organic solution was separated andthe aqueous solution was extracted with EtOAc (2×40 mL). The organicsolutions were combined, washed with brine, dried over MgSO₄, filteredand concentrated to give 4-(6-methyl-5-nitropyridin-2-yl)morpholine (4.0g, 99%), as a muddy green solid. LCMS (FA): R_(t)=1.59 min, m/z=224.0(M+H).

Step 2: N-(2-Methyl-6-morpholin-4-ylpyridin-3-yl)guanidine

The hydrochloride salt ofN-(2-methyl-6-morpholin-4-ylpyridin-3-yl)guanidine was prepared from4-(6-methyl-5-nitropyridin-2-yl)morpholine following the procedures ofGeneral Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-{6-[4-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}guanidineN-{2-methyl-6-[methyl(1-methyl-pyrrolidin-3-yl)amino]pyridin-3-yl}guanidineN-{2-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}guanidineN-(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)guanidineN-{2-methyl-6-[(1-methylpiperidin-4-yl)oxy]pyridin-3-yl}guanidineN-(2-methyl-6-piperidin-1-ylpyridin-3-yl)guanidineN-(2-methyl-6-pyrrolidin-1-ylpyridin-3-yl)guanidineN-[2-methyl-6-(4-methylpiperazin-1-yl)pyridin-3-yl]guanidineN-[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]guanidineN-[6-(isopropylamino)-2-methylpyridin-3-yl]guanidineN-[2-methyl-6-(tetrahydro-2H-pyran-4-ylamino)pyridin-3-yl]guanidineN-{2-methyl-6-[methyl(1-methyl-piperidin-4-yl)amino]pyridin-3-yl}guanidineN-(6-{[2-(dimethylamino)ethyl]amino}-2-methylpyridin-3-yl)guanidineN-{6-[[2-(dimethylamino)ethyl](methyl)amino]-2-methyl-pyridin-3-yl}guanidineN-{6-[[3-(dimethylamino)propyl](methyl)amino]-2-methyl-pyridin-3-yl}guanidineN-(6-{[3-(dimethylamino)propyl]amino}-2-methylpyridin-3-yl)guanidineN-{6-[3-(dimethylamino)piperidin-1-yl]-2-methylpyridin-3-yl}guanidineN-[2-methyl-6-(piperidin-4-ylamino)pyridin-3-yl]guanidineN-(2-methyl-6-{[(1-methyl-piperidin-4-yl)methyl]amino}pyridin-3-yl)guanidineN-{4-methyl-6-[(1-methylpiperidin-4-yl)amino]pyridin-3-yl}guanidine

N-[6-(Dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]guanidine

Step 1: 1,6-Dimethyl-3,5-dinitropyridin-2(1H)-one

To a solution of 1,6-dimethylpyridin-2(1H)-one (3.40 g, 27.60 mmol) inacetic anhydride (35 mL) was added fuming nitric acid (15 mL) dropwiseover 3 h. The reaction mixture was allowed to stir for an additional 3h, then the reaction mixture was cooled to 0° C. and water (30 mL) wasadded. The mixture was filtered and the solid was purified by columnchromatography to give 1,6-dimethyl-3,5-dinitropyridin-2(1H)-one (1.82g, 30%).

Step 2: 1,6-tert-Butyl(2-methyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-yl)carbamate

To a sealed tube was added 1,6-dimethyl-3,5-dinitropyridin-2(1H)-one(0.16 g, 0.76 mmol), tert-butyl (4-oxocyclohexyl)carbamate (0.19 g, 0.91mmol), and ammonia (1M in methanol, 9 mL). The resulting mixture wasallowed to stir for 5 h at 60° C. and then allowed to cool to rt andconcentrated. The residue was purified by column chromatography to givetert-butyl (2-methyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-yl)carbamate(0.066 g, 28%) as a solid.

Step 3: N,N-2-Trimethyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-amine

To a round bottom flask was added tert-butyl(2-methyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-yl)carbamate (0.14 g,0.46 mmol) and HCl (4M in dioxane, 8 mL). The resulting mixture wasallowed to stir for 5 h. The mixture was concentrated, and then water(10 mL), sodium triacetoxyborohydride (0.29 g, 1.38 mmol), andformaldehyde (37% in water, 6 mL) were added. The resulting mixture wasallowed to stir for 80 min. Solid NaOH was added until the solution wasbasic. The mixture was extracted with EtOAc (2×30 mL). The organicsolutions were combined, dried over Na₂SO₄, filtered and concentrated togive N,N-2-trimethyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-amine (0.11 g,97%) as a solid.

Step 4:N-[6-(Dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]guanidine

The hydrochloride salt ofN-[6-(dimethylamino)-2-methyl-5,6,7,8-tetrahydroquinolin-3-yl]guanidinewas prepared fromN,N-2-trimethyl-3-nitro-5,6,7,8-tetrahydroquinolin-6-amine following theprocedures of General Method 1. LCMS (FA): R_(t)=0.24 min, m/z=248.2(M+H).

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-(2,6-dimethyl-5,6,7,8-tetrahydro-1,6-naphthyridin-3-yl)guanidine

N-{5-[2-(Dimethylamino)ethyl]-2-methylpyridin-3-yl}guanidine

Step 1: 2-Methyl-3-nitro-5-[(trimethylsilyl)ethynyl]pyridine

To a solution of 5-bromo-2-methyl-3-nitropyridine (9.38 g, 43.2 mmol)and (trimethylsilyl)acetylene (9.16 mL, 64.8 mmol) in triethylamine (300mL) were added bis(triphenylphosphine)palladium(II) chloride (3.34 g,4.75 mmol) and copper(I) iodide (1.64 g, 8.64 mmol). The reactionmixture was allowed to stir for 4 h at 80° C. and then cooled to rt andfiltered through Celite®. The filter pad was washed with EtOAc and thefiltrate was washed with saturated aqueous NaHCO₃ (500 mL) then brine(500 mL). The organic solution was dried over Mg₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to give2-methyl-3-nitro-5-[(trimethylsilyl)ethynyl]pyridine (9.22 g, 91%). LCMS(FAL): R_(t)=11.04 min, m/z=235.1 (M+H).

Step 2: 5-Ethynyl-2-methyl-3-nitropyridine

A solution of 2-methyl-3-nitro-5-[(trimethylsilyl)ethynyl]pyridine (2.32g, 9.90 mmol) in methanol (22 mL) was cooled to 0° C. and potassiumcarbonate (137 mg, 0.99 mmol) was added. The reaction mixture wasallowed to warm and stir at rt for 10 min, during which time a yellowprecipitate formed. The reaction mixture was then concentrated anddiluted with ether (200 mL). The organic solution was washed with water(200 mL) and brine (200 mL), dried over MgSO₄, filtered and concentratedto give 5-ethynyl-2-methyl-3-nitropyridine (1.48 g, 92%) as a solid.LCMS (FA): R_(t)=1.64 min, m/z=163.0 (M+H).

Step 3: N,N-Dimethyl-2-(6-methyl-5-nitropyridin-3-yl)ethanamine

To a solution of 5-ethynyl-2-methyl-3-nitropyridine (930 mg, 5.74 mmol)in EtOH (7 mL) were added dimethylamine hydrochloride (2.34 g, 28.7mmol) and sodium cyanoborohydride (0.721 g, 11.5 mmol). The reactionmixture was allowed to stir at reflux for 24 h, concentrated, andtreated with 4N NaOH (10 mL). The solution was extracted with DCM (2×200mL). The organic solutions were combined, washed with brine (200 mL),dried over MgSO₄, filtered and concentrated. The residue was purified bycolumn chromatography to giveN,N-dimethyl-2-(6-methyl-5-nitropyridin-3-yl)ethanamine (489 mg, 41%).LCMS (AA): R_(t)=1.35 min, m/z=210 (M+H).

Step 4: N-{5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}guanidine

The hydrochloride salt ofN-{5-[2-(dimethylamino)ethyl]-2-methylpyridin-3-yl}guanidine wasprepared from N,N-dimethyl-2-(6-methyl-5-nitropyridin-3-yl)ethanaminefollowing the procedures of General Method 1.

Compounds in the following table were prepared as the hydrochloride saltfrom the appropriate starting materials using the procedures describedabove:

N-[2-methyl-5-(2-piperidin-1-ylethyl)pyridin-3-yl]guanidineN-[2-methyl-5-(2-morpholin-4-ylethyl)pyridin-3-yl]guanidineN-(5-{2-[3-(dimethylamino)piperidin-1-yl]ethyl}-2-methyl-pyridin-3-yl)guanidineN-[2-methyl-5-(2-pyrrolidin-1-ylethyl)pyridin-3-yl]guanidineN-(5-{2-[4-(dimethylamino)piperidin-1-yl]ethyl}-2-methyl-pyridin-3-yl)guanidine

N-{2-Methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}guanidine

Step 1: 2-Methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-amine

To a solution of 5-ethynyl-2-methyl-3-nitropyridine (600 mg, 3.70 mmol)in EtOH (10 mL) was added 1-methyl-piperazine (1.64 mL, 14.8 mmol). Thereaction mixture was allowed to stir at reflux overnight. The reactionmixture was allowed to cool to rt and palladium hydroxide on carbon(0.060 g) was added to the solution. The reaction mixture was purgedwith hydrogen and then allowed to stir under an atmosphere of H₂ at rtovernight. The reaction mixture was filtered through celite and thefilatrate was concentrated. The residue was purified by columnchromatography to give2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-amine (0.664 g,77%).

Step 2:N-{2-Methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}guanidine

The hydrochloride salt ofN-{2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-yl}guanidinewas prepared from2-methyl-5-[2-(4-methylpiperazin-1-yl)ethyl]pyridin-3-amine followingthe procedures of General Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-{5-[2-(cyclopentylamino)ethyl]-2-methylpyridin-3-yl}guanidineN-{5-[2-(2,6-dimethylmorpholin-4-yl)ethyl]-2-methyl-pyridin-3-yl}guanidineN-(5-{2-[3-(dimethylamino)pyrrolidin-1-yl]ethyl}-2-methyl-pyridin-3-yl)guanidine

tert-Butyl[3-(5-{[amino(imino)methyl]amino}-6-methylpyridin-3-yl)propyl]carbamate

Step 1: tert-Butyl[3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-yl]carbamate

To a solution of 5-bromo-2-methyl-3-nitropyridine (2.81 g, 12.9 mmol) inTEA (25 mL) was added tert-butyl prop-2-yn-1-ylcarbamate (2.58 g, 16.6mmol). The solution was degassed with argon. To the reaction mixturewere added bis(triphenylphosphine)palladium(II) chloride (0.18 g, 0.26mmol) and copper(I) iodide (0.098 g, 0.52 mmol). The reaction mixturewas allowed to stir at 70° C. for 2.5 h and was then concentrated anddiluted with EtOAc (100 mL). The mixture was filtered over a pad ofCelite® and the resulting filtrate was washed with sat. aq. NaHCO₃ (100mL). The organic solution was dried over MgSO₄, filtered andconcentrated. The residue was purified by column chromatography to givetert-butyl [3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-yl]carbamate(2.98 g, 79%) as a yellow solid. LCMS (FA): R_(t)=1.81 min, m/z=292.0(M+H).

Step 2: tert-Butyl [3-(5-amino-6-methylpyridin-3-yl)propyl]carbamate

To a solution of tert-butyl[3-(6-methyl-5-nitropyridin-3-yl)prop-2-yn-1-yl]carbamate (2.98 g, 10.2mmol) in MeOH (100 mL) was added palladium hydroxide on carbon (0.72 g).The reaction vessel was flushed with H₂ and then the reaction mixturewas allowed to stir at rt under an atmosphere of hydrogen overnight. Thereaction mixture was filtered over a pad of Celite® and the filtrate wasconcentrated to give tert-butyl[3-(5-amino-6-methylpyridin-3-yl)propyl]carbamate (2.97 g, >99%) as ayellow oil. LCMS (FA): R_(t)=0.85 min, m/z=266.0 (M+H).

Step 3: Dibenzyl{(Z)-[(5-{3-[(tert-butoxycarbonypamino]propyl}-2-methylpyridin-3-yl)amino]methylylidene}biscarbamate

To a solution of tert-butyl[3-(5-amino-6-methylpyridin-3-yl)propyl]carbamate (2.99 g, 11.3 mmol),1,3,-bis(benzyloxycarbonyl)-2-methyl-2-thiopseudourea (3.41 g, 9.51mmol), and mercury(II) chloride (3.37 g, 12.4 mmol) in DCM (100 mL) wasadded TEA (5.19 mL, 37.3 mmol) dropwise via syringe. The reactionmixture was allowed to stir at rt overnight. The reaction mixture wasfiltered over a pad of Celite® and the resulting filtrate wasconcentrated. The residue was purified by column chromatography to givedibenzyl{(Z)-[(5-{3-[(tert-butoxycarbonyl)amino]propyl}-2-methylpyridin-3-yl)amino]methylylidene}biscarbamate(3.34 g, 61%) as a white solid. LCMS (FA): R_(t)=1.90 min, m/z=576.0(M+H).

Step 4: tert-butyl[3-(5-{[amino(imino)methyl]amino}-6-methylpyridin-3-yl)propyl]carbamate

To a solution of dibenzyl{(Z)-[(5-{3-[(tert-butoxycarbonyl)amino]propyl}-2-methylpyridin-3-yl)amino]methylylidene}biscarbamate(3.34 g, 5.80 mmol) in MeOH (100 mL) was added palladium hydroxide oncarbon (0.41 g). The reaction vessel was flushed with hydrogen gas andthen allowed to stir at rt under an atmosphere of hydrogen gasovernight. The reaction mixture was filtered over a pad of Celite® andthe filtrate was concentrated to give tert-butyl[3-(5-{[amino(imino)methyl]amino}-6-methylpyridin-3-yl)propyl]carbamate(1.77 g, 5.76 mmol, 99%) as a white solid. LCMS (FA): R_(t)=0.76 min,m/z=308.0 (M+H).

N-{5-[3-(Dimethylamino)propyl]-2-fluoropyridin-3-yl}guanidine

Step 1: 5-[3-(Dimethylamino)prop-1-yn-1-yl]-2-fluoropyridin-3-amine

A mixture of 5-bromo-2-fluoropyridin-3-amine (1.00 g, 5.24 mmol),N,N-dimethylprop-2-yn-1-amine (0.68 mL, 6.28 mmol), and cesium carbonate(3.41 g, 10.5 mmol) in DMF (25 mL) was degassed with argon for 15 min.To this slurry were added bis(acetonitrile)palladium(II) chloride (0.068g, 0.26 mmol) and Xphos (0.324 g, 0.68 mmol). The suspension was allowedto stir at 50° C. for 4 h under an atmosphere of nitrogen and was thenconcentrated. The residue was purified by column chromatography to give5-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluoropyridin-3-amine (0.46 g,45%).

Step 2: N-{5-[3-(Dimethylamino)propyl]-2-fluoropyridin-3-yl}guanidine

The hydrochloride salt ofN-{5-[3-(dimethylamino)propyl]-2-fluoropyridin-3-yl}guanidine wasprepared from5-[3-(dimethylamino)prop-1-yn-1-yl]-2-fluoropyridin-3-amine followingthe procedures of General Method 1.

N-{5-[3-(Dimethylamino)propyl]pyridin-3-yl}guanidine

Step 1: 5-[3-(Dimethylamino)prop-1-yn-1-yl]pyridin-3-amine

To a degassed solution of 5-bromopyridin-3-amine (1.00 g, 5.78 mmol) andN,N-dimethylprop-2-yn-1-amine (1.57 mL, 14.6 mmol) in TEA (26 mL) wereadded bis(triphenylphosphine)palladium(II) chloride (0.203 g, 0.29 mmol)and copper iodide (0.110 g, 0.58 mmol). The reaction mixture was allowedto stir under an atmosphere of argon at 80° C. for 14 h. The reactionmixture was allowed to cool to rt and then filtered through Celite. Thefilter pad was washed with EtOAc and the filtrate was concentrated. Theresidue was purified by column chromatography to give5-[3-(dimethylamino)prop-1-yn-1-yl]pyridin-3-amine (0.55 g, 54%).

Step 2: N-{5-[3-(Dimethylamino)propyl]pyridin-3-yl}guanidine

The hydrochloride salt ofN-{5-[3-(dimethylamino)propyl]pyridin-3-yl}guanidine was prepared from5-[3-(dimethylamino)prop-1-yn-1-yl]pyridin-3-amine following theprocedures of General Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-{5-[4-(dimethylamino)butyl]-2-methylpyridin-3-yl}guanidine

N-(2,4-Dimethylpyridin-3-yl)guanidine

The hydrochloride salt of N-(2,4-dimethylpyridin-3-yl)guanidine wasprepared from 2,4-dimethyl-3-nitropyridine following the procedures ofGeneral Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-(2,6-dimethylpyridin-3-yl)guanidine

5-{[Amino(imino)methyl]amino}-6-methyl-N-(2-morpholin-4-ylethyl)pyridine-2-carboxamide

Step 1: 6-Methyl-N-(2-morpholin-4-ylethyl)-5-nitropyridine-2-carboxamide

To a solution of 6-methyl-5-nitropyridine-2-carboxylic acid (0.500 g,3.0 mmol) in DCM (7 mL) were added 2-morpholin-4-ylethanamine (1.0 mL,7.7 mmol), TBTU (0.992 g, 3.1 mmol), and DMA (1.34 mL, 7.7 mmol). Thereaction mixture was allowed to stir at rt overnight and then quenchedwith water. The organic solution was separated and the aqueous solutionwas extracted with DCM. The organic solutions were combined andconcentrated. The residue was purified by column chromatography to give6-methyl-N-(2-morpholin-4-ylethyl)-5-nitropyridine-2-carboxamide (0.27g, 30%).

Step 2:5-{[Amino(imino)methyl]amino}-6-methyl-N-(2-morpholin-4-ylethyl)pyridine-2-carboxamide

The hydrochloride salt of5-{[amino(imino)methyl]amino}-6-methyl-N-(2-morpholin-4-ylethyl)pyridine-2-carboxamidewas prepared from6-methyl-N-(2-morpholin-4-ylethyl)-5-nitropyridine-2-carboxamide fromthe appropriate starting materials following the procedures of GeneralMethod 1.

N-[2-Methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]guanidine

Step 1: 1-Methyl-4-(6-methyl-5-nitropyridin-3-yl)piperazine

1-Methylpiperazine (1.79 mL, 16.1 mmol) was added to a suspension of5-bromo-2-methyl-3-nitropyridine (1.00 g, 4.61 mmol),4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (267 mg, 0.461 mmol),tris(dibenzylideneacetone)dipalladium (0) (170 mg, 0.180 mmol) andcesium carbonate (3.00 g, 9.22 mmol) in 1,4-dioxane (20.0 mL) underargon in a sealed reaction vessel. The reaction mixture was allowed tostir for 22 h, at 150° C. The reaction mixture was next partitionedbetween EtOAc (50 mL) and water (50 mL). The organic solution wasseparated and the aqueous solution was extracted with EtOAc (3×50 mL).The organic solutions were combined, washed with brine (twice), driedover MgSO₄, filtered and concentrated. The residue was purified bycolumn chromatography to give1-methyl-4-(6-methyl-5-nitropyridin-3-yl)piperazine (0.77 g, 71%) as abrown solid. LCMS (FA): R_(t)=1.07 min, m/z=237.3 (M+H).

Step 2: N-[2-Methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]guanidine

The hydrochloride salt ofN-[2-methyl-5-(4-methylpiperazin-1-yl)pyridin-3-yl]guanidine wasprepared from 1-methyl-4-(6-methyl-5-nitropyridin-3-yl)piperazinefollowing the procedures of General Method 1.

Compounds in the following table were prepared as the hydrochloridesalts from the appropriate starting materials using the proceduresdescribed above:

N-{5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methyl-pyridin-3-yl}guanidineN-{5-[(3R,5S)-4-ethyl-3,5-dimethylpiperazin-1-yl]-2-methyl-pyridin-3-yl}guanidineN-{5-[(3S)-4-ethyl-3-methylpiperazin-1-yl]-2-methyl-pyridin-3-yl}guanidine

Example 9 Synthesis of Pyridyl Halides Used for the Preparation ofCompounds of Formula vi3-(5-Bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropan-1-amine

Step 1: (2E)-3-(5-Bromo-2-methoxypyridin-3-yl)-N,N-dimethylacrylamide

To a solution of (2E)-3-(5-bromo-2-methoxypyridin-3-yl)acrylic acid(1.00 g, 3.87 mmol) in DCM (10 mL) were added dimethylamine (2.0 M inTHF, 5.80 mmol), DIEA (2.02 mL, 11.6 mmol) and TBTU (1.49 g, 4.65 mmol).The reaction mixture was allowed to stir at rt overnight. The reactionwas quenched with water and then the aqueous solution was extracted withDCM. The organic solutions were combined, washed with brine, dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography to give(2E)-3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylacrylamide (1.05 g,95%) as a white solid. LCMS (FA): R_(t)=1.68 min, m/z=285.1 (M+H).

Step 2: 3-(5-Bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropanamide

To a solution of(2E)-3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylacryamide (1.05 g,3.68 mmol) in EtOH (50 mL) was added platinum on sulfided carbon (5% Pt,600 mg). The reaction mixture was flushed with hydrogen gas and allowedto stir at rt for 1 h under an atmosphere of hydrogen. The reactionmixture was filtered through a Celite® pad and washed with MeOH. Thefiltrate was concentrated to give3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropanamide (1.0 g, 94%)as an oil. LCMS (FA): R_(t)=1.64 min, m/z=287.2 (M+H).

Step 3: 3-(5-Bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropan-1-amine

To a solution of3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropanamide (1.0 g, 3.0mmol) in THF (40 mL) was added borane-tetrahydrofuran (1.0 M in THF,8.71 mL) slowly. The reaction mixture was allowed to stir for 3 h at 50°C. The mixture was allowed to cool to rt and then concentrated todryness. To the residue, a solution of HCl (3.0 M in water, 40 mL) wasadded. The resulting mixture was allowed to stir for 2 h at 80° C. Themixture was allowed to cool to rt and then was basified to pH>10 with apotassium hydroxide solution. The aqueous solution was extracted withDCM. The organic solutions were combined, washed with brine, dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography to give3-(5-bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropan-1-amine (0.377 g,40%) as an oil. LCMS (FA): R_(t)=1.03 min, m/z=273.1 (M+H).

3-(5-Bromo-2-methoxypyridin-3-yl)-N,N-dimethylpropan-1-amine

Step 1: 2-Iodo-3,5-dibromopyridine

To a sealed tube was added 2,3,5-tribromopyridine (5.63 g, 17.83 mol),sodium iodide (8.02 g, 53.50 mol), propanenitrile (45 mL), andchlorotrimethylsilane (2.26 mL, 17.83 mol). The resulting mixture wasallowed to stir for 50 min at 105° C. The mixture was allowed to cool tort and 2M NaOH (50 mL) was added. The mixture was extracted with EtOAc(2×80 mL). The organic solutions were combined, dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography to give 2-iodo-3,5-dibromopyridine (5.4 g, 83%) as asolid.

Step 2: 3,5-Dibromo-2-(trifluoromethyl)pyridine

To a solution of 2-iodo-3,5-dibromopyridine (4.21 g, 11.60 mmol) in NMP(25 mL) were added copper(I) iodide (4.42 g, 23.21 mmol), potassiumfluoride (1.35 g, 23.21 mmol), and (trifluoromethyl)trimethylsilane(5.61 mL, 35.90 mmol). The resulting mixture was allowed to stirovernight at 50° C. The mixture was poured into 12% aqueous ammonia, andthen extracted with Et₂O (2×120 mL). The organic solutions werecombined, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography to give3,5-dibromo-2-(trifluoromethyl)pyridine (1.63 g, 46%) as a liquid.

Step 3: 3-Bromo-5-[(E)-2-ethoxyvinyl]-2-(trifluoromethyl)pyridine

To a solution of 3,5-dibromo-2-(trifluoromethyl)pyridine (1.11 g, 3.65mmol) in water (9 mL) were added2-[(E)-2-ethoxyvinyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.72 g,3.65 mmol), tetrakis(triphenylphosphine)-palladium(0) (0.42 g, 0.36mmol), 1,2-dimethoxyethane (16.7 mL), and sodium carbonate (1.16 g, 11.0mmol). The resulting mixture was allowed to stir overnight at 60° C.After the reaction was allowed to cool to rt, EtOAc (50 mL) was added.The organic solution was separated, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to give3-bromo-5-[(E)-2-ethoxyvinyl]-2-(trifluoromethyl)pyridine (0.41 g, 38%).

Step 4:2-[5-Bromo-6-(trifluoromethyl)pyridin-3-yl]-N,N-dimethylethanamine

To a solution of3-bromo-5-[(E)-2-ethoxyvinyl]-2-(trifluoromethyl)-pyridine (0.35 g, 1.18mmol) in AcOH (9 mL) was added water (3 mL). The resulting mixture wasallowed to stir overnight at 105° C. The mixture was allowed to cool tort. Dimethylamine hydrochloride (0.96 g, 11.8 mmol) and DCM (40 mL) wereadded. The resulting mixture was allowed to stir for 2 h. Sodiumtriacetoxyborohydride (0.50 g, 2.36 mmol) was added in portions over 50min. The resulting mixture was allowed to stir for 3 h and was thenconcentrated. To the residue was added saturated NaHCO₃ (40 mL), and themixture was extracted with EtOAc (2×30 mL). The organic solutions werecombined, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography to give2-[5-bromo-6-(trifluoromethyl)pyridin-3-yl]-N,N-dimethylethanamine(0.052 g, 15%) as a solid. LCMS (AA): R_(t)=1.24 min, m/z=297.0 (M+H).

4-[2-(5-Bromopyridin-3-yl)ethyl]morpholine

Step 1: 4-[(5-Bromopyridin-3-yl)acetyl]morpholine

To a solution of (5-bromopyridin-3-yl)acetic acid (1.00 g, 4.63 mmol)and morpholine (0.50 mL, 5.73 mmol) in DCM (50 mL) was added EDC (1.78g, 9.30 mmol) and 1-hydroxybenzotriazole (1.25 g, 9.26 mmol). Thereaction mixture was allowed to stir at rt for 50 min and then dilutedwith DCM (50 mL) and 1N HCl (100 mL). The organic solution was separatedand discarded. The aqueous solution was neutralized with sat. aq. NaHCO₃(100 mL) and was extracted with DCM (3×50 mL). The organic solutionswere combined, dried over MgSO₄, filtered and concentrated to give4-[(5-bromopyridin-3-yl)acetyl]morpholine (1.26 g, 4.40 mmol, 95%) as alight brown solid. LCMS (FA): R_(t)=1.11 min, m/z=287.0 (M+H).

Step 2: 4-[2-(5-Bromopyridin-3-yl)ethyl]morpholine

To a solution of 4-[(5-bromopyridin-3-yl)acetyl]morpholine (1.26 g, 4.40mmol) in THF (15 mL) was added a 1M solution of borane in THF (23.2 mL,23.2 mmol). The reaction mixture was allowed to stir at 55° C. overnightand then was allowed to cool to rt and then diluted with water (100 mL)and EtOAc (100 mL). The organic solution was separated and the aqueoussolution was extracted with EtOAc (2×50 mL). The organic solutions werecombined, dried over MgSO₄, filtered and concentrated to give a yellowoil, which was dissolved in 1N HCl (100 mL). This solution was allowedto stir at 80° C. overnight. The reaction mixture was neutralized withsat. aq. NaHCO₃ (100 mL) and then extracted with EtOAc (3×50 mL). Thecombined organic solutions were washed with brine, dried over MgSO₄,filtered and concentrated to give4-[2-(5-bromopyridin-3-yl)ethyl]morpholine (0.631 g, 2.33 mmol, 53%) asa light yellow oil. LCMS (FA): R_(t)=0.50 min, m/z=273.0 (M+H).

Compounds in the following table were prepared from the appropriatestarting materials using the procedures described above:

2-(5-bromopyridin-3-yl)-N,N-dimethylethanamine1-[2-(5-bromopyridin-3-yl)ethyl]-4-methylpiperazine

5-Bromo-2-(pyrrolidin-1-yl)pyridine

A solution of 2,5-dibromopyridine (1.60 g, 6.76 mmol) and pyrrolidine(5.64 mL, 67.6 mol) in 1-butanol (10.0 mL) was allowed to stir in asealed reaction vessel while heating at 75° C. for 16 h. The reactionsolvent was evaporated to produce an oily solid which was purified bycolumn chromatography to give 5-bromo-2-(pyrrolidin-1-yl)pyridine (1.50g, 6.60 mmol, 98%) as a white solid. LCMS (FA): m/z=229.1 (M+H).

Compounds in the following table were prepared from the appropriatestarting materials using the procedure described above:

1-(5-bromopyridin-2-yl)-4-methylpiperazine4-(5-bromopyridin-2-yl)morpholineN-(5-bromopyridin-2-yl)-N,N′,N′-trimethylpropane-1,3-diamine

N-(5-Chloropyridin-2-yl)-2,2-dimethylpropanamide

TEA (6.78 mL, 48.6 mmol) was added to a suspension of2-amino-5-chloropyridine (5.00 g, 38.9 mmol) in DCM (75 mL). Theresulting solution was cooled to 0° C. and a solution of2,2-dimethylpropanoyl chloride (5.74 mL, 46.7 mmol) in DCM (10.0 mL) wasadded dropwise over a 5 min period. The resulting mixture was allowed tostir at rt for 1 h. The reaction mixture was then transferred to aseparatory funnel and washed with a saturated aqueous solution ofNaHCO₃. The organic solution was dried over MgSO₄, filtered andconcentrated. The residue was purified by column chromatography to giveN-(5-chloropyridin-2-yl)-2,2-dimethylpropanamide (8.0 g, 40 mmol, 99%).LCMS (FA): m/z=215.1 (M+H).

3-(5-Bromo-2,6-dimethylpyridin-3-yl)-N,N-dimethylprop-2-yn-1-amine

To a mixture of 3,5-dibromo-2,6-dimethylpyridine (605 mg, 2.54 mmol),copper iodide (22 mg, 0.11 mmol) andbis(triphenylphosphine)palladium(II) chloride (41 mg, 0.058 mmol) indiethylamine (3.0 mL) was added propargyl(dimethylamine) (0.27 mL, 2.50mmol). The reaction mixture was allowed to stir for 15 h at 50° C. andwas then allowed to cool to rt. The resulting mixture was diluted withEtOAc (30 mL) and 1M aqueous Na₂CO₃ (20 mL). After stirring for 30 min,the organic solution was separated and the aqueous solution wasextracted with EtOAc (2×15 mL). The organic solutions were combined,washed with water and brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography to give3-(5-bromo-2,6-dimethylpyridin-3-yl)-N,N-dimethylprop-2-yn-1-amine (313mg, 51%) as a dark oil. LCMS (AA): R_(t)=1.47 min, m/z=267.1 (M−H),269.1 (M+H).

1-(5-Bromopyridin-3-yl)-4-methylpiperazine

To a suspension of 3,5-dibromopyridine (0.30 g, 4.6 mmol),4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (57 mg, 0.098 mmol),tris(dibenzylideneacetone)dipalladium (0) (32 mg, 0.035 mmol) and cesiumcarbonate (830 mg, 2.5 mmol) in 1,4-dioxane (4.0 mL) in a sealed rxnvessel was added 1-methylpiperazine (0.170 mL, 1.5 mmol) under argon.The reaction mixture was heated for 30 min in a microwave reactor, at150° C. The reaction mixture was diluted with EtOAc (50 mL), washed withbrine and water, dried over Na₂SO₄, then filtered and concentrated. Thecrude reaction product was purified by column chromatography to give1-(5-bromopyridin-3-yl)-4-methylpiperazine (0.13 g, 39%) as a brown oil.LCMS (FA): R_(t)=1.03 min, m/z=258.0 (M+H).

Compounds in the following table were prepared from the appropriatestarting materials using the procedure described above:

(2R)-4-(5-bromopyridin-3-yl)-1-ethyl-2-methylpiperazine(2S)-4-(5-bromopyridin-3-yl)-1-ethyl-2-methylpiperazine

Example 10 Synthesis of2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneMonohydrochloride Salt

A mixture of2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(131.0 g, 269 mmol) in 200P ethanol (1.05 L) was heated to reflux. Asolution formed. Hydrochloric acid (1.25 M in ethanol, 43 mL, 53.8 mmol)was added slowly. Seeds were added. More hydrochloric acid (1.25 M inethanol, 172 mL, 215 mmol) was added dropwise over 4 hours. Thesuspension formed was kept at reflux for 1 hour, then allowed to cooldown to room temperature in 5 hours. The product was collected byfiltration then washed with EtOH (20 mL). Drying under vacuo at 35° C.overnight afforded2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneMonohydrochloride Salt as a light yellow solid (128 g, HPLC purity >99%,90.9% yield). LCMS (FA): R_(t)=6.08 min, m/z=485.2 (M−H). FTIR: 1575,1434, 1408, 1334, 1315, 1173, 1136, 1116, 1087, 787 cm⁻¹.

Example 11 Synthesis of2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneDihydrochloride Salt

To a solution of2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thione(153 mg, 0.314 mmol) in Ethanol (5.1 mL, 87 mmol) under gentle reflux(oil bath at 85° C.) was added 0.89 M of Hydrochloric acid in Ethanol(7.0E2 μL, 0.63 mmol) (850 μl of conc. HCl in 10 ml of ethanoltritrated). The resulting solution was seeded and shortly thereaftersolid formed. The mixture was allowed to cool at room temperature in theoil bath and was left overnight at room temperature. The solid wasfiltered off and dried under high vacuum to afford2-({5-[3-(Dimethylamino)propyl]-2-methylpyridin-3-yl}amino)-9-(trifluoromethyl)-5,7-dihydro-6H-pyrimido[5,4-d][1]benzazepine-6-thioneDihydrochloride Salt as a yellow solid (163 mg, 92.6% yield).

Expression and Purification of Protein Kinase Enzymes Example 12 PLK1Enzyme Expression and Purification

Recombinant human PLK1 was expressed in E. coli as an N-terminal Smtfusion protein using a proprietary vector (pSGX4) by Structural Genomics(SGX). The fusion partner was removed through cleavage with Ulp1 afteran initial purification using a Ni2+ affinity column.

Example 13 Protein Kinase Enzyme Assays

PLK1 DELFIA® Kinase Assay

The human PLK1 enzymatic reaction totaled 30 μL contained 50 mM Tris-HCl(pH 8.0), 10 mM MgCl₂, 0.02% BSA, 10% glycerol, 1 mM DTT, 100 mM NaCl,3.3% DMSO, 50 μM ATP, 2 μM peptide substrate(Biotin-AHX-LDETGHLDSSGLQEVHLA-CONH₂) and 0.3 nM recombinant humanPLK1[2-369]T210D. The enzymatic reaction mixture, with or without PLKinhibitors, was incubated 90 minutes at room temperature beforetermination with 50 μL of STOP buffer containing 1% BSA, 0.05% Tween20,100 mM EDTA. 50 μL of the stopped enzyme reaction mixture wastransferred to a Neutravidin-coated 384-well plate (Pierce) andincubated at room temperature for 60 minutes. The wells were washed withwash buffer (25 mM Tris, 3 mM KCl, 14 mM NaCl and 0.1% Tween-20) andincubated for 1 hour with 50 μL of antibody reaction mixture containing1% BSA, 0.05% Tween-20, anti-phospho-cdc25c rabbit monoclonal antibody(325 μM, Millennium Pharmaceuticals), europium labeled anti-rabbit IgG(2 nM, Perkin Elmer) in DELPHIA assay buffer (Perkin Elmer). The wellswere washed and then the bound europium was liberated using 50 μL ofEnhancement Solution (Perkin Elmer). Quantification of europium was doneusing a Pherastar (BMG Labtech)

The following compounds have PLK1 Delfia IC₅₀<5 nM:

I-1, I-2, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12, I-13, I-14,I-15, I-17, I-18, I-19, I-21, I-22, I-23, I-24, I-25, I-26, I-27, I-28,I-29, I-31, I-32, I-34, I-38, I-39, I-40, I-42, I-43, I-45, I-46, I-47,I-49, I-50, I-51, I-52, I-53, I-54, I-55, I-57, I-58, I-59, I-60, I-61,I-62, I-63, I-64, I-65, I-66, I-67, I-68, I-69, I-72, I-73, I-74, I-75,I-76, I-77, I-78, I-80, I-81, I-84, I-85, I-86, I-87, I-88, I-89, I-90,I-91, I-92, I-94, I-95, I-96, I-99, I-100, I-103, I-104, I-105, I-106,I-107, I-108, I-109, I-111, I-113, I-114, I-115, I-119

The following compounds have PLK1 Delfia IC₅₀ 5-25 nM:

I-3, I-16, I-33, I-35, I-36, I-41, I-44, I-56, I-70, I-71, I-79, I-82,I-83, I-93, I-97, I-101, I-102, I-110, I-112, I-116, I-117, I-118,I-121, I-122, I-123

The following compounds have PLK1 Delfia IC₅₀>25 nM:

I-30, I-37, I-98, I-120

Example 14 Cellular Assays

PLK1 Phosphorylation Assay

Inhibition of PLK1 activity in whole cell systems can be assessed bydetermination of decreased phosphorylation of PLK1 substrates. Forexample, determining decreased phosphorylation of cdc25c on Threonine96, a PLK1 substrate can be used to measure inhibition of PLK1 activityin a whole cell system. Alternatively, any known PLK1 substrate can beused in similar assay methods to assess inhibition of PLK1 activity.

In a specific example, HeLa cells were seeded in a 96-well cell cultureplate (4.5×10³ cells/well) in Minimum Essential Medium supplemented with10% FBS and incubated overnight at 37° C., 5% CO₂. pSGcdc25C (0.05 ug)and pcDNA3.1 PLK T210D (0.02 ug) DNA were transfected using 0.15 μlFugene 6 (Roche) transfection reagent in each well. Cells were incubatedwith PLK1 inhibitors for 2 hours at 37° C., fixed with 4%paraformaldehyde for 15 minutes and then permeabilized for 15 minuteswith 0.5% TritonX-100 in PBS. 100 μl of Roche blocking buffer was addedto the wells prior to incubation with rabbit anti-pcdc25c T96 (1:2500,Millennium Pharmaceuticals) and mouse anti-myc (clone 9E10) (1:250,Millennium Pharmaceuticals Inc.) antibodies overnight at 4° C. Afterwashing with PBS the cells were stained with anti-rabbit IgG Alexa 488(1:500, Molecular Probes) and anti-mouse IgG Alexa 660 (1:500) for 45minutes at room temperature. DNA was then stained with Hoechst solution(2 μg/ml). The percentage of pcdc25c and anti-myc positive cells werequantified using the Opera instrument and Acapella Image Analysis(Perkin Elmer.)

Mitotic Index Assay

HT29 cells (2.5×10³ cells/well) in 75 μl of McCoy's 5A media(Invitrogen) supplemented with 10% FBS (Invitrogen) were seeded in wellsof a 96-well Optilux plate (BD bioscience) and incubated for 24 hours at37° C., 5% CO₂. 25 μl of serially diluted test compounds in McCoy's 5Amedia supplemented with 10% FCS (Qibco) were added to the wells. Cellswere incubated for 24 hrs at 37° C. and then fixed with the addition of50 μl of 4% paraformaldehyde for 10 minutes and permeabilized for 10minutes with the addition of 50 μl 0.5% TritonX-100 in PBS. Afterwashing with PBST, 50 μl of 0.5% Roche blocking buffer was added to thewells and incubated for 1 hour at room temperature. Cells were thenincubated with mouse anti-pH is H3 (1:500, Millennium Pharmaceuticals)for 1 hour at room temperature. After washing with PBST the cells werestained with anti-mouse IgG Alexa 594 (1:200) for 1 hour at roomtemperature. DNA was then stained with Hoechst solution (2 μg/ml). Thepercentage of pHisH3 cells were quantified using Discovery-1 andMetaMorph (Molecular Devices.)

Anti-Proliferation Assays

8 μl of serially diluted test compounds were added to 75 μl of HT29(2.66×10⁴ cells/ml) cells in McCoy's 5A media supplemented with 10% FBS(Invitrogen) in Biocoat Poly-D lysine 384 well Black/Clear plates (BDBiosciences). Cells were incubated for 72 hrs at 37° C., 5% CO₂.Supernatant was aspirated from the wells, leaving 25 μl in each well.ATP-lite 1 step reagent (25 μl, Perkin Elmer) was added to each well andluminescence for each plate was read on the LeadSeeker (AmershamBiosciences). Percent inhibition was calculated using the values from aDMSO control set to 100%.

Example 15 In Vivo Assays

In Vivo Tumor Efficacy

HT29 human colon cancer cells with p53 deficiency are cultured inMcCoy's 3A medium containing 10% FCS and incubated at 37° C., 5% CO₂.The cells are trypsinized and resuspended in Hanks buffer at 2×10⁷cells/mL. 100 μL of the cell suspension (2×10⁶ cells) is asepticallyinjected into the subcutaneous space in the right dorsal flank of femaleNCR nude mice (age 7-10 weeks, Taconic) using a 23-ga needle. Seven daysafter implantation, the tumors are measured in two dimensions (lengthand width) with a caliper and the animal body weight is measured with abalance. Tumor volume is calculated with the following formula: tumorvolume=L×W²×0.5. When the average tumor volume reaches about 200 mm³,the individual animals are assigned to different study groups using arandom number generation method. The typical study consists of vehiclecontrol, PLK1 inhibitor alone groups at various doses and schedules (10animals/group). Tumor size and body weight are measured twice a week forthree to four weeks. Once the tumor volume reaches over 10% of the bodyweight of the animal, or the mouse body weight loss is more than 20%,the mouse is euthanized. Data is collected and only those study groupswhich end the study with at least 7 animals are used for the analysis.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments, which utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments, which have been represented by way of example.

1. A compound of formula IV:

or a salt thereof, wherein R¹ is selected from hydrogen, —CN, halogen,optionally substituted C₁₋₆aliphatic, or —YR^(1a), wherein Y is —O—,—S—, or —NR^(1a), and each occurrence of R^(1a) is independentlyhydrogen, or optionally substituted C₁₋₆aliphatic; R² is selected fromhydrogen, halogen, —ZR^(2a), or —OR^(2b), wherein Z is an optionallysubstituted C₁₋₆ alkylene chain, and R^(2a) is —OR^(2b), —N(R^(2b))₂,—SR^(2b), —C(O)N(R^(2b))₂, —N(R^(2b))C(O)R^(2b), —SO₂N(R^(2b))₂,—NR^(2b)SO₂R^(2b), —NR^(2b)C(O)N(R^(2b))₂, or —NR^(2b)SO₂N(R^(2b))₂,wherein each occurrence of R^(2b) is independently hydrogen oroptionally substituted C₁₋₆alkyl, or two occurrences of R^(2b), takentogether with a nitrogen atom to which they are bound, form anoptionally substituted 3-7-membered heterocyclyl ring; R³ is selectedfrom hydrogen, halogen, optionally substituted C₁₋₄-alkyl, or optionallysubstituted C₁₋₄-alkoxy; R⁴ is selected from hydrogen, optionallysubstituted C₁₋₆aliphatic, an optionally substituted 3-7-memberedheterocyclyl ring, —(CH₂)_(x)NR^(4a)R^(4b), —(CH₂)_(x)NR^(4a)C(O)R^(4b),—(CH₂)_(x)NR^(4a)S(O)₂R^(4b), —(CH₂)_(x)C(O)R^(4b),—(CH₂)_(x)C(O)NR^(4a)R^(4b), —(CH₂)_(x)S(O)₂NR^(4a)R^(4b), or—(CH₂)_(x)OR^(4b), wherein each occurrence of x is independently 0-6;wherein R^(4a) is hydrogen or optionally substituted C₁₋₆aliphatic, andR^(4b) is hydrogen, optionally substituted C₁₋₆aliphatic, optionallysubstituted C₃₋₇heterocyclyl or C₃₋₇carbocyclyl ring, or is W—R^(4c),wherein W is an optionally substituted C₂₋₆ alkylene chain, and R^(4c)is an optionally substituted C₃₋₇-heterocyclyl ring, —OR^(4d),—N(R^(4d))₂, —SR^(4d), —C(O)N(R^(4d))₂, —N(R^(4d))C(O)R^(4d),—SO₂N(R^(4d))₂, —NR^(4d)SO₂R^(4d), —NR^(4d)C(O)N(R^(4d))₂, or—NR^(4d)SO₂N(R^(4d))₂, wherein each occurrence of R^(4d) isindependently hydrogen or optionally substituted C₁₋₆aliphatic, or twooccurrences of R^(4d), taken together with the nitrogen atom to whichthey are bound, form an optionally substituted 3-7-membered heterocyclylring; or wherein R^(4a) and R^(4b), taken together with the nitrogenatom to which they are bound, form an optionally substituted3-7-membered heterocyclyl ring; R⁵ is hydrogen, optionally substitutedC₁₋₆aliphatic, an optionally substituted C₃₋₇heterocyclyl ring, or isX—R^(5a), wherein X is an optionally substituted C₂₋₆ alkylene chain or—NR^(5c), wherein when X is an optionally substituted C₂₋₆alkylene chainR^(5a) is —OR^(5b), —N(R^(5b))₂, —SR^(5b), —C(O)N(R^(5b))₂,—N(R^(5b))C(O)R^(5b), —SO₂N(R^(5b))₂, —NR^(5b)SO₂R^(5b),—NR^(5b)C(O)N(R^(5b))₂, or —NR^(5b)SO₂N(R^(5b))₂; and when X is—NR^(5c), R^(5a) is hydrogen or optionally substituted C₁₋₆aliphatic, orR^(5a) and R^(5c), taken together with the nitrogen atom to which theyare bound, form an optionally substituted 3-7-membered heterocyclylring; wherein each occurrence of R^(5b) and R^(5c) is independentlyhydrogen or optionally substituted C₁₋₆aliphatic, or two occurrences ofR^(5b), or R^(5a) and R^(5c), taken together with the nitrogen atom towhich they are bound, form an optionally substituted 3-7-memberedheterocyclyl ring; or wherein R⁴ and R⁵, taken together, form anoptionally substituted 5-7-membered cycloaliphatic or heterocyclyl ring;and R⁶ is selected from hydrogen, halogen, optionally substitutedC₁₋₄alkyl, or optionally substituted C₁₋₄-alkoxy.